Efficacy of high-intensity laser therapy in patients with adhesive capsulitis: a sham-controlled randomized controlled trial.
Adhesive capsulitis is an idiopathic condition characterized by painful shoulder stiffness, impairs quality of life, and causes disability. The purpose of the study is to evaluate the effects of high-intensity laser therapy (HILT) on pain, disability, and quality of life in patients with adhesive capsulitis. The study was designed as a prospective, double-blinded, and sham-controlled randomized trial. Thirty-six patients diagnosed with adhesive capsulitis were randomized into HILT plus therapeutic exercises, sham-laser plus therapeutic exercises, and control-therapeutic exercises only groups. All groups received 25 min of exercises to the shoulder joint supervised by a physiotherapist. Patients in both the HILT and the sham-laser group were blinded to their group randomization. The interventions were performed five times a week for 3 weeks (a total of 15 sessions). The primary outcome measure was the visual analogue scale for pain (Visual Analog Scale for pain (VAS)-pain). The secondary outcome measures were shoulder pain and disability index (SPADI), short-form health survey-36 (SF-36), and range of motion (ROM) of the shoulder joint. Assessments were performed at pre-intervention, post-intervention, and 12-week follow-up by an investigator who was blinded. Thirty-one participants completed the study. HILT plus therapeutic exercises showed significant differences in VAS-pain, SPADI-pain, and SF-36 (subgroups of energy/fatigue, pain, and general health) scores (all P < 0.05). All groups provided comparable improvements in SPADI-disability and ROM of shoulder joint after the interventions and during the follow-up (all P < 0.05). Fifteen sessions of HILT are superior to improve pain and quality of life but not superior in terms of disability or function in patients with adhesive capsulitis. Clinical trial registration number: The study was registered at the US National Institutes of Health ( ClinicalTrials.gov ) (NCT03929276) and available at https://clinicaltrials.gov/ct2/show/NCT03929276?term=tu%C4%9Fba+atan&draw=2&rank=3.


Infectivity assays for the Autographa californica nuclear polyhedrosis virus using Spodoptera littoralis cells.
Tissue culture ID50 and plaque assays for the Autographa californica nuclear polyhedrosis virus, using the Spodoptera littoralis cell line HPB-SL, were developed. Direct comparison using these assay methods showed that these cells were as susceptible to infection as the more commonly used Spodoptera frugiperda cell line IPLB-SF-21. Both infectious tissue culture supernatants or virus isolated directly from polyhedra could be titrated. It was important to use low cell seeding densities in the assays so that clear centres of infection formed. Dose-response curves indicated that one infectious particle was capable of initiating an infection. Virus could be cloned using either method even though, for the plaque assay, plates had been stained. The tissue culture ID50 assay was performed using 96-well plates and required an incubation period of about 10 days. The plaque assay used a simple nutrient agarose overlay and an incubation period of 5-6 days. Easily countable plaques of 0.3-1.2 mm diameter were detected after staining with iodonitrote-trazolium chloride. The plaques comprised areas of inhibited cell division and round or dead cells. Most plaques contained only some cells with polyhedra and yields averaged about 1/cell. Occasionally plaques or infected wells were found in which no polyhedra could be seen. These infectivity assays are therefore not dependent on polyhedra formation.


Plasma anti-Müllerian hormone as a biomarker for bovine granulosa-theca cell tumors: comparison with immunoreactive inhibin and ovarian steroid concentrations.
Granulosa-theca cell tumors (GTCTs) are the most frequently reported ovarian tumors in cattle. Clinically, GTCTs could be confused with other ovarian abnormalities; therefore, the only definitive diagnosis for such tumors is histopathology of a biopsy from the affected ovary. However, this is an invasive technique and unsuitable for farm conditions. As a result, the key aim of this study was to evaluate the diagnostic value of anti-Müllerian hormone (AMH), a glycoprotein hormone that is synthesized exclusively by ovarian granulosa cells, as a sensitive noninvasive biomarker for diagnosing GTCTs in cattle. To achieve this aim, we conducted two experiments. In experiment 1, four clinically healthy Japanese Black cows had blood samples taken daily over one estrous cycle to characterize their AMH profiles throughout the estrous cycle. Additionally, single blood samples were collected from 21 cyclic cows to clarify the physiological range of AMH. In experiment 2, cows with histologically confirmed GTCT (GTCT group, n = 9) and cows affected with cystic ovarian disease (COD group, n = 8) had one blood sample taken before extraction of the tumorous ovary or therapeutic treatment for the COD. Blood samples (n = 105) from cyclic cows (n = 25) in experiment 1 were assigned as a physiologically cyclic group (PC group). Plasma AMH, immunoreactive inhibin (ir-INH), estradiol-17β (E2), testosterone (T), and progesterone (P4) concentrations were assayed in all samples. In experiment 1, the mean plasma AMH concentration was 0.09 ± 0.003 ng/mL and did not show substantial fluctuation throughout the estrous cycle. In experiment 2, plasma AMH, ir-INH, and E2 concentrations were significantly elevated in the GTCT group in comparison with the PC group; among these parameters, only the AMH concentrations were significantly higher in the GTCT group than in the COD group. The area under the receiver operating characteristic curve of AMH for diagnosis of GTCT was 0.99 and was significantly higher than that of ir-INH (P < 0.001) and E2 (P < 0.01). Moreover, the AMH at a cutoff point of ≥0.36 ng/mL had the highest diagnostic accuracy (99.2%), sensitivity (100%), and specificity (99.1%) compared with the other tested parameters. In conclusion, plasma AMH concentration is probably a more reliable and sensitive biomarker for bovine GTCTs than the concentrations of ir-INH or ovarian steroids.


[Mechanism of lymphocyte activation by periodic acids. Blastogenesis and induction of immunomodulator populations].
Mild periodate treatment is mitogenic for T lymphocytes. With murine spleen cells periodate oxidation is effective between C8 and C9 on sialyl acid residues. With human blood lymphocytes this oxidation occurs between C7 and C8 of these residues. In vitro immune response is inhibited by periodate treatment. Activation of an immunosuppressive T lymphocyte population is obtained. Similar results are performed with human blood lymphocytes.


Metabolism of 6,9,12-octadecatrienoic acid and 6,9,12,15-octadecatetraenoic acid by rat hepatocytes.
When 5.10(6) hepatocytes were incubated for 40 min with 0.015-0.3 mM (1-14C)-labeled 6,9,12-octadecatrienoic acid or (1-14C)-labeled 6,9,12,15-octadecatetraenoic acid there was a concentration-dependent acylation of radioactive metabolites into both phospholipids and triacylglycerol. However, when the concentration of either substrate exceeded 60-150 microM there was no further increase in the metabolism of either substrate to longer-chain (n-6) and (n-3) acids. When cells were then incubated for various periods of time with 60 microM substrate there was initial rapid removal of the substrate which was accompanied by its acylation into lipids. Over time, the amount of both substrates in lipids declined without an overall drop in specific activity. This decline was accompanied by an increase in long-chain (n-6) and (n-3) fatty acids. Similar results were obtained when the time-dependent metabolism of the two substrates was examined in individual hepatocyte phospholipids. Collectively, these findings suggest that when these two 18-carbon acids are produced by desaturation of dietary linoleate and linolenate that they are in part initially acylated into a labile phospholipid pool. Rapid release and subsequent further metabolism to longer-chain (n-6) and (n-3) acids may explain why these products of the 6-desaturase do not accumulate in membrane lipids.


Interactions of the subthalamic nucleus and the subpallidal area in oro-facial dyskinesia: role of GABA and glutamate.
Previous studies have shown that lowering the GABAergic activity in the sub-pallidal area (SP) in the cat results in the display of oro-facial dyskinesia (OFD). There exists an intense, mutual anatomical connection between the SP and the subthalamic nucleus and the adjoining lateral hypothalamic area (STH). The present study investigated whether the STH is also involved in OFD. Once this turned out to be true (see below), it was investigated whether the SP-specific OFD is funneled via the STH, or vice versa. Bilateral injections of low doses (50-250 ng) of picrotoxin, a non-competitive GABA antagonist, into the STH were found to elicit OFD. This effect which was quantified in terms of numbers of tongue protrusions, was dose-dependent: a bell-shaped dose-response was found (50-500 ng). The OFD elicited by the most effective dose of picrotoxin (250 ng) was significantly antagonized by muscimol, a specific GABAA agonist, in a dose (50 ng) which itself was ineffective, indicating GABA specificity. In addition, it was found that OFD elicited by local injections of picrotoxin (250 ng) into the STH was significantly attenuated by SP injections of the broad spectrum glutamate antagonist kynurenic acid in a dose (1000 ng) which itself was ineffective, but not by muscimol (100 ng), indicating that the STH-elicited OFD needs an intact and functioning glutaminergic, but not GABAergic, transmission process in the SP for its expression.(ABSTRACT TRUNCATED AT 250 WORDS)


Exploring epitranscriptomics for crop improvement and environmental stress tolerance.
Climate change and stressful environmental conditions severely hamper crop growth, development and yield. Plants respond to environmental perturbations, through their plasticity provided by key-genes, governed at post-/transcriptional levels. Gene-regulation in plants is a multilevel process controlled by diverse cellular entities that includes transcription factors (TF), epigenetic regulators and non-coding RNAs beside others. There are successful studies confirming the role of epigenetic modifications (DNA-methylation/histone-modifications) in gene expression. Recent years have witnessed emergence of a highly specialized field the "Epitranscriptomics". Epitranscriptomics deals with investigating post-transcriptional RNA chemical-modifications present across the life forms that change structural, functional and biological characters of RNA. However, deeper insights on of epitranscriptomic modifications, with >140 types known so far, are to be understood fully. Researchers have identified epitranscriptome marks (writers, erasers and readers) and mapped the site-specific RNA modifications (m6A, m5C, 3' uridylation, etc.) responsible for fine-tuning gene expression in plants. Simultaneous advancement in sequencing platforms, upgraded bioinformatic tools and pipelines along with conventional labelled techniques have further given a statistical picture of these epitranscriptomic modifications leading to their potential applicability in crop improvement and developing climate-smart crops. We present herein the insights on epitranscriptomic machinery in plants and how epitranscriptome and epitranscriptomic modifications underlying plant growth, development and environmental stress responses/adaptations. Third-generation sequencing technology, advanced bioinformatics tools and databases being used in plant epitranscriptomics are also discussed. Emphasis is given on potential exploration of epitranscriptome engineering for crop-improvement and developing environmental stress tolerant plants covering current status, challenges and future directions.


Ratings of perceived exertion in individuals with varying fitness levels during walking and running.
It was the purpose of this investigation to: 1) compare the ratings of perceived exertion (RPEs) in high and low fit individuals when walking and running at comparable exercise intensities and 2) to determine if ventilation (VE) provides a central signal for RPEs. Nine high fit and nine low fit male subjects completed two exercise bouts on a treadmill, one uphill walking and the other level running. Workloads for each bout were set at 90% of each subject's ventilatory threshold (VT) as determined from a graded exercise test. Oxygen consumption (Vo2), heart rate (HR), and VE were all similar between the walk and run trials for the low fit subjects (P greater than 0.05). HR were found to be significantly greater during the walk trial vs. the run trial (P less than 0.05) for the high fit subjects, whereas, VE was significantly greater during the run trial. Oxygen consumption was similar for the high fit subjects during both trials (P greater than 0.05). During the walk and run trials, central (12.1 +/- 1.6 vs. 11.4 +/- 1.5), local (14.0 +/- 1.3 vs. 13.9 +/- 1.1) and overall (12.8 +/- 1.2 vs. 12.4 +/- 1.4) RPEs were not found to be significantly different for the low fit group (P greater than 0.05). In contrast, during the walk vs. the run trial there was a significant increase in central (10.7 +/- 2.0 vs. 9.2 +/- 1.9), local (11.5 +/- 2.0 vs. 9.8 +/- 1.8) and overall (11.2 +/- 2.4 vs. 9.6 +/- 2.3) RPEs for the high fit group (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Regulation of [3H]norepinephrine release by N-methyl-D-aspartate receptors in minislices from the dentate gyrus and the CA1-CA3 area of the rat hippocampus.
It has been reported previously that N-methyl-D-aspartic acid induces a significant release of [3H]norepinephrine preaccumulated in slices from the hippocampus. In the present study, we investigated whether there are regional differences in the hippocampus regarding this N-methyl-D-aspartate effect. In the absence of Mg2+, N-methyl-D-aspartate (10-200 microM) induced the release of [3H]norepinephrine from superfused minislices containing the dentate gyrus area or the CA1-CA3 region of the hippocampus. Such N-methyl-D-aspartate effects on [3H]norepinephrine release were significantly higher in the dentate gyrus than in the CA1-CA3 area. The N-methyl-D-aspartate effects in both hippocampal areas were also reduced significantly by D-2-amino-5-phosphonovaleric acid (50 microM), an antagonist of the N-methyl-D-aspartate receptor, and by tetrodotoxin, a blocker of the voltage-dependent Na+ channels. The extent of this reduction was the same in the dentate gyrus and the CA1-CA3 area. Further experiments, conducted in the presence of Mg2+, demonstrated that N-methyl-D-aspartic acid increased K(+)-induced release of [3H]norepinephrine from dentate gyrus minislices but not from the CA1-CA3 area. The results are consistent with the existence of a higher density and/or different subtypes of N-methyl-D-aspartate receptors modulating [3H]norepinephrine release in the dentate gyrus as compared with the CA1-CA3 hippocampal area.


Acetazolamide in the treatment of pyruvate dysmetabolism syndromes.
Two children with periodic weakness beginning in infancy were demonstrated to have abnormalities in pyruvate metabolism by the production of lactic acidosis following a glucose load. Daily oral doses of acetazolamide reduced the frequency of attacks and reversed the abnormal response to glucose loading. The mechanism of action of acetazolamide in these patients is not clear.


Characterising sex differences of autosomal DNA methylation in whole blood using the Illumina EPIC array.
Sex differences are known to play a role in disease aetiology, progression and outcome. Previous studies have revealed autosomal epigenetic differences between males and females in some tissues, including differences in DNA methylation patterns. Here, we report for the first time an analysis of autosomal sex differences in DNAme using the Illumina EPIC array in human whole blood by performing a discovery (n = 1171) and validation (n = 2471) analysis. We identified and validated 396 sex-associated differentially methylated CpG sites (saDMPs) with the majority found to be female-biased CpGs (74%). These saDMP's are enriched in CpG islands and CpG shores and located preferentially at 5'UTRs, 3'UTRs and enhancers. Additionally, we identified 266 significant sex-associated differentially methylated regions overlapping genes, which have previously been shown to exhibit epigenetic sex differences, and novel genes. Transcription factor binding site enrichment revealed enrichment of transcription factors related to critical developmental processes and sex determination such as SRY and ESR1. Our study reports a reliable catalogue of sex-associated CpG sites and elucidates several characteristics of these sites using large-scale discovery and validation data sets. This resource will benefit future studies aiming to investigate sex specific epigenetic signatures and further our understanding of the role of DNA methylation in sex differences in human whole blood.


Effect of antibodies to human seminal plasma inhibin on spermatogenesis and sperm agglutination in adult male rats.
In vitro incubation of rat epididymal sperm with antiserum to human seminal plasma inhibin (As hSPI) caused agglutination of the sperm. In vivo administration of As hSPI to adult male rats resulted in a significant decrease in testicular as well as epididymal sperm counts. Furthermore, the majority (almost 90%) of the epididymal sperm were agglutinated. When these animals were mated with normal cycling females, significant reduction in fertility was observed.


Microvascular responses to endothelin in deoxycorticosterone acetate-salt hypertensive rats.
The objective of the present study was to determine if endothelin-1 played a role in the elevated peripheral resistance in deoxycorticosterone-acetate (DOCA)-salt hypertension. Radioimmunoassay showed that the concentration of endothelin-1 was higher in thoracic aorta of the DOCA-salt group than that of the control normotensive (CN) group. Responses of arterioles in the rat cremaster to endothelin-1 were also observed using in vivo closed circuit television microscopy. Microvascular sensitivity to endothelin-1 was decreased in the DOCA-salt group. In the presence of an endothelin type A (ET-A) receptor antagonist, low concentrations of endothelin-1 induced a significant vasoconstriction in the DOCA-salt group. In the presence of endothelin type B (ET-B) receptor antagonist, microvascular responses to endothelin-1 were attenuated in the DOCA-salt group. These results indicated that the increased tissue level of endothelin-1 may decrease the ET-A receptor-mediated vascular response to endothelin-1. However, the ET-B receptor-mediated vasoconstriction is potentiated during DOCA-salt hypertension.


Early Feasibility of Endovascular Repair of Distal Aortic Arch Aneurysms Using Patient-Specific Single Retrograde Left Subclavian Artery Branch Stent Graft.
To describe the feasibility and outcomes of endovascular repair of distal aortic arch aneurysms using a patient-specific stent graft with a pre-loaded single retrograde left subclavian artery (LSA) branch stent graft. We reviewed the clinical data and outcomes of consecutive patients enrolled in an ongoing prospective, non-randomized physician-sponsored investigational device exemption study to evaluate the outcomes of endovascular aortic arch repair using patient-specific arch branch stent grafts (William Cook Europe, Bjaeverskov, Denmark) between 2019 and 2022. All patients received a design with triple-wide scallop and a single retrograde LSA branch with a pre-loaded catheter. There were five male patients with median age of 77 years old (72-80) treated using the single LSA branch stent graft. Technical success was achieved in all patients. Median operating time, fluoroscopy time, and total radiation dose area product were 103 (78-134) minutes, 26 (19-39) minutes, and 123 (71-270) mGy.cm2, respectively. There were no 30-day or in-hospital mortality, neurological or other major adverse events (MAEs). During median follow-up of 21 (20-27) months, all patients were alive with patent LSA branches, except for one who died of COVID-19 complications. There was no branch instability or secondary interventions. This early feasibility study demonstrates successful endovascular repair of distal aortic arch aneurysms using a patient-specific stent graft with single retrograde LSA branch without technical failures, mortality or neurological events. Larger clinical experience and longer follow-up are needed to determined effectiveness of this approach in patients who need endovascular repair with proximal extension into Zone 2.


Urine catecholamine excretion after large doses of fentanyl, fentanyl and diazepam and fentanyl, diazepam and pancuronium.
The effects of fentanyl (0.5 mg/kg iv), fentanyl with diazepam (1 mg/kg iv) and fentanyl, diazepam and pancuronium (0.1 mg/kg iv) on heart rate (HR), mean arterial blood pressure (BP), cardiac output (QT), urine flow rate and urine epinephrine and norepinephrine excretion were determined in nine dogs. Fentanyl did not significantly change QT or BP but did reduce HR and urine flow rate (P less than 0.05). Urine epinephrine and norepinephrine excretion rates were signicantly increased by fentanyl (P less than 0.05). Diazepam caused no significant further changes in QT, BP or HR 30 minutes after administration, but urine epinephrine and norepinephrine excretion rates were reduced to control (pre-fentanyl) levels. Addition of pancuronium after fentanyl and diazepam increased urine flow rate to pre-fentanyl levels and elevated QT, BP and HR above controls but produced no significant change in urine epinephrine or norepinephrine excretion. These data suggest that fentanyl increases catecholamine blood levels and imply that the latter may be one mechanism by which cardiovascular dynamics are maintained stable during fentanyl anaesthesia. Our findings also demonstrate that cardiovascular stimulation after pancuronium is not associated with increased urinary catecholamine excretion.


Physiological responses to digestion in low salinity in the crabs Carcinus maenas and Cancer irroratus.
Osmoregulation and digestion are energetically demanding, and crabs that move into low salinity environments to feed must be able to balance the demands of both processes. Achieving this balance may pose greater challenges for weak than for efficient osmoregulators. This study examined the rate of oxygen consumption (MO2) of Carcinus maenas (efficient osmoregulator) and Cancer irroratus (weak osmoregulator) as a function of feeding and hyposaline stress. The MO2 increased 2-fold in both species following feeding. The MO2 increased and remained elevated in fasted crabs during acute hyposaline exposure. When hyposaline stress occurred after feeding, C. maenas responded with an immediate summation of the MO2 associated with feeding and hyposaline stress, whereas C. irroratus reacted with a partial summation of responses in a salinity of 24‰, but were unable to sum responses in 16‰. C. irroratus exhibited longer gut transit times. This may be due to an inability to regulate osmotic water onload as efficiently as C. maenas. Mechanical digestion in crabs can account for a significant portion of SDA, and a short term interruption led to the delay in summation of metabolic demands. Although protein synthesis is reported to account for the majority of SDA, this did not appear to be the case here. Protein synthesis rates were higher in C. irroratus but neither feeding or salinity affected protein synthesis rates of either species which suggests that protein synthesis can continue in low salinity as long as substrates are available.


[Epidemiological study of periodontal disease. Mass examination in university students with CPITN].
The purpose of this study was to investigate the prevalence and treatment needs of periodontal disease in the 1,150 university students (male: 726, female: 424) by using simplified Community Periodontal Index of Treatment Needs (CPITN-S). The results were as follows: 1. Approximately 85% of all the subjects have something wrong in periodontal tissue. If examined in detail; 1) Code 4: a group (1.6%) who had a deep pocket more than 6 mm. 2) Code 3: a group (20.1%) who had a shallow pocket 4 to 5 mm. 3) Code 2: a group (55.9%) who had supra- or subgingival calculus. 4) Code 1: a group (7.5%) who had bleeding on probing only. 2. As for treatment needs (TN), approximately 78% of the subjects need to receive professional prophylaxis and more complex treatment including periodontal surgery (TN 2-3). 3. Average time required to examine one subject was about one minute, so we could cut down the measuring time. From this point, we have understood that simplified CPITN was greatly effective to curtail the time for the primary screening of periodontal disease in the young age group.


The biology of VSIG4: Implications for the treatment of immune-mediated inflammatory diseases and cancer.
V-set and immunoglobulin domain containing 4 (VSIG4), a type I transmembrane receptor exclusively expressed in a subset of tissue-resident macrophages, plays a pivotal role in clearing C3-opsonized pathogens and their byproducts from the circulation. VSIG4 maintains immune homeostasis by suppressing the activation of complement pathways or T cells and inducing regulatory T-cell differentiation, thereby inhibiting the development of immune-mediated inflammatory diseases but enhancing cancer progression. Consequently, VSIG4 exhibits a potential therapeutic effect for immune-mediated inflammatory diseases, but also is regarded as a novel target of immune checkpoint inhibition in cancer therapy. Recently, soluble VSIG4, the extracellular domain of VSIG4, shed from the surface of macrophages, has been found to be a biomarker to define macrophage activation-related diseases. This review mainly summarizes recent new findings of VSIG4 in macrophage phagocytosis and immune homeostasis, and discusses its potential diagnostic and therapeutic usage in infection, inflammation, and cancer.


Risk factors for distant metastases in head and neck squamous cell carcinoma.
To evaluate the frequency of distant metastases (DM) and to determine the ability of certain clinical and pathologic factors to predict the development of distant metastases. Retrospective analysis. University hospital. A total of 1972 patients with oral, oropharyngeal, hypopharyngeal, and laryngeal squamous cell carcinomas who were treated from 1981 to 1998 and who were not diagnosed as having DM at the time of initial treatment. We evaluated the frequency of DM and the influence of different variables in their appearance. A total of 181 patients (9.2%) (95% confidence interval, 7.9%-10.5%) developed DM. Younger age (<45 years), hypopharyngeal localization, an advanced T stage and/or N stage tumor according to the TNM staging system, high histologic grade, and locoregional control were found to be significantly associated with the risk of DM at both univariate and multivariate analyses. The incidence of DM in subjects with head and neck squamous cell carcinoma is relatively low. The risk of DM is influenced by age, site of primary cancer, local and/or regional extension, grading, and achievement of locoregional control.


Immunochemical characterization of anti-acetylcholine receptor antibodies in primary biliary cirrhosis.
Although the presence of anti-mitochondrial antibodies is the main characteristic of primary biliary cirrhosis (PBC), other autoantibodies have been described in this disease. This study employs immunoblot methods to test whether the sera of PBC patients also contain antibodies directed against nicotinic acetylcholine receptors (AChR). We show that the majority of patients' sera indeed react with AChR just as sera of myasthenic patients do. In contrast, however, these anti-AChR antibodies do not lead to significant clinical symptoms of myasthenia. In all cases studied, PBC sera recognized a protein with the molecular weight of the alpha-chain of acetylcholine receptor (40 kDa). In addition, with both liver mitochondria and AChR-rich membranes as antigens, PBC sera reacted with proteins with apparent molecular weights around 68 kDa and the same pI values. This protein is not present in purified AChR preparations. These data suggest structural, if not functional, relationships between membrane components of liver mitochondria and muscle endplates.


Role of calcium and crossbridges in modulation of rates of force development and relaxation in skinned muscle fibers.
The influence of Ca2+ and force generating crossbridges on the kinetics of force development and relaxation was examined in skinned muscle fibers activated by photolytic release of Ca2+ from a caged calcium or inactivated by photolytic uptake of Ca2+ by a caged Ca2+ chelator. In frog fibers at 10 degrees C, decreasing the Ca2+ released from caged calcium to an extent that resulted in 50% of maximum force development produced an approximate seven-fold decrease in the rate of contraction. In contrast decreasing the number of force generating crossbridges by partial extraction of troponin C (TnC) or addition of vanadate caused only minor changes in contraction rate. Thus the rate of force development decreases dramatically with decreases in the Ca2+ level which suggests that a step in the crossbridge cycle may be Ca2+ dependent. The kinetics of relaxation induced by photolysis of diazo-2 was: a) slowed by stabilization of the sarcomeres by repeated releases and re-stretches during contraction and b) accelerated when the amplitude of force development was decreased by decreasing the [Ca2+] which induced a steady contraction. The half time of relaxation decreased by approximately two- to three-fold, when 50% of maximum force was developed. One interpretation of these results is that decreasing the number of force generating crossbridges may speed relaxation by inducing a decreased affinity of TnC for Ca2+ and thus accelerating the Ca2+ dissociation rate from TnC and thereby increasing relaxation rate.


Long-term adherence to home mechanical ventilation: a 10-year retrospective, single-centre cohort study.
Sleep-disordered breathing (SBD) can be associated with hypercapnic respiratory failure (HRF). Home Mechanical Ventilation (HMV) is the preferred long-term treatment for patients with chronic HRF. We reviewed the database of a large tertiary referral centre for HMV to study the long-term adherence to HMV in chronic hypercapnic patients. Data on adherence and characteristics of patients who received HMV for the treatment of SDB were collected over a decade using electronic patient records. The primary outcome parameter in this study was annual non-adherence rate (patients with HMV usage of <4 hours/night in the service divided by the number of all new patients of the same year), secondary outcomes were patients' characteristics and reasons for low adherence. HMV adherence clinics were established to improve uptake. Two thousand and two hundred twenty-eight patients with HRF were under active follow-up on HMV at the end of the recording period. In contrast, a total of 1,900 patients had their HMV contracts terminated over the course of a decade (due to non-adherence, transfer to other services or death). Out of those, 222 patients {62 [52-72] years, body-mass index, BMI 40 [35-43] kg/m2, 58.1% male, Epworth Sleepiness Scale, ESS 9 [4-15] points, 4% oxygen desaturation index, 4%ODI 32 [20-71] × hour-1, TcCO2 6.6 [6.0-7.2] kPa} met the non-adherence criteria (nocturnal usage 0-4 hours). The annual non-adherence rate was 25.5% of all new setups in 2010, and declined to 3.4% in 2019 (relative reduction of 86%, P<0.001). Patients with Obstructive Sleep Apnoea/Obesity Hypoventilation Syndrome (58.2%), Neuromuscular Diseases (NMD) (26.8%) and COPD (13.6%) accounted for most cases of this non-adherent cohort. The vast majority of the patients (96.1%) were established on full-face masks. In 23.4% of patients, substantial weight loss (>10%) was the most common reason for low adherence; general displeasure (21.3%), uncontrolled symptoms (12.8%), claustrophobia (6.7%), mood (4.8%) and mask intolerance (4.3%) caused problems as well. Non-adherence to HMV in patients with chronic HRF can affect significant proportions of patients. However, the non-adherent rate substantially decreases when individual treatment solutions are offered in multi-disciplinary clinics.


Unilateral paralysis of the diaphragm during pregnancy: case report and literature review.
Unilateral diaphragmatic paralysis is usually asymptomatic and diagnosed incidentally and often does not need any additional diagnostic workup, but in the setting of pregnancy it may require additional investigations. We describe a patient in her last trimester of pregnancy (36th week) who presented to the hospital with symptoms suggestive of upper respiratory tract infection and with a chest radiograph showing left diaphragmatic eventration, for which she underwent thoracotomy and cesarean section. In the report, we review the normal respiratory changes associated with pregnancy and discuss the management of patients with diaphragmatic paralysis and pregnancy.


Second Primary Ovarian Epithelial, Fallopian Tube, and Primary Peritoneal Cancers after Breast Cancer Diagnosis: Korea Central Cancer Registry.
A prior history of breast cancer is a risk factor for the subsequent development of primary peritoneal, epithelial ovarian, and fallopian tubal (POFT) cancers. This study aimed to estimate the incidence of secondary POFT malignancy in breast cancer patients and the clinical outcomes of primary and secondary POFT cancer. We searched the Korea Central Cancer Registry to find patients with primary and secondary POFT cancer who had breast cancer in 1999-2017. The incidence rate and standardized incidence ratio were calculated. Additionally, we compared the overall survival of patients with primary and secondary POFT cancer. Based on the age-standardized rate, the incidence of second primary POFT cancer after breast cancer was 0.0763 per 100,000 women, which increased in Korea between 1999 and 2017. Among the 30,366 POFT cancer patients, 25,721 were primary POFT cancer only, and 493 had secondary POFT cancer after a breast cancer diagnosis. Second primary POFT cancer patients were older at the time of diagnosis (55 vs. 53, p < 0.001) and had a larger proportion of serous histology (68.4% vs. 51.2%, p < 0.001) than patients with primary POFT. There were no differences between the two groups in tumor stage at diagnosis. The 5-year overall survival rates were 60.2% and 56.3% for primary and secondary POFT cancer, respectively (p=0.216). The incidence of second primary POFT cancer after breast cancer increased in Korea between 1999 and 2017. Besides, second primary POFT cancer patients were diagnosed at older ages and had more serous histology.


Readers' forum: experiences with a system. The implementation of a completely operational computer system in a cardio-thoracic department.
Wythenshawe Hospital is a large general district hospital with more than 1000 beds, situated approximately 9 miles south of Manchester. The Cardio-Thoracic department, housed in the new section of the hospital which was completed in 1973, is the major centre for cardiothoracic surgery in the greater Manchester area. It also receives any major accident cases which require treatment within the Intensive Therapy Unit. This paper describes a Patient Data Display System and the methods used to introduce the concept of such a system, to doctors, nurses, laboratory and paramedical staff, and the manner in which these staff were trained in the use of computer terminals. Principles underlying the need for and requirements of such a system are also discussed. Six months after hardware commissioning the system was fully operational throughout the whole department.


An acrylic ampoule represents a denture surface in a microcalorimetric biofilm problem.
An acrylic microcalorimetric ampoule with inserts represented a denture surface in a Streptococcus mutants biofilm study. The biofilm of micro-organisms did not contain as many micro-organisms as those previously studied in planktonic suspension. A more sensitive microcalorimeter (Thermometric's thermal activity monitor) was required to detect the metabolism of S. mutants adhered to an engineered acrylic flow-through ampoule. Biofilm heat outputs were obtained in the acrylic ampoule following standardized procedures with cryopreserved aliquots of a homogeneous suspension of S. mutants.


Anaerobic co-digestion of three commercial bio-plastic bags with food waste: Effects on methane production and microbial community structure.
The emergence of bioplastic bags as a replacement for traditional petroleum-based plastic bags is promising for their simultaneous anaerobic digestion with food waste. In this study, the degradation of three bioplastic bags is evaluated during anaerobic co-digestion with food waste under mesophilic/thermophilic conditions, and the results indicated PBAT/PLA/starch > PLA > PBAT for methane production rate. The PBAT/PLA/starch mixture produced 23.4 ml/g of methane at 55 °C, and the cumulative methane production increased by 28.4 % compared to the control. In addition, the lag time before methane production was reduced by one to four days when anaerobic co-digestion was performed under thermophilic conditions, and the conversion of the bioplastics improved by 9.11-11.2 %. Microscopy further showed obvious physical degradation of the PBAT/PLA/starch material. The FTIR analysis showed that the characteristic peaks of the material at 3320, 2957, and 934 cm-1 decreased significantly after anaerobic fermentation. The biodegradability of the polymer decreased with an increase in the content of the crystalline area in the structure. The addition of a comonomer reduced the crystallinity of the polymer. In addition, the biodegradability was increased by adjusting the hydrolysis reaction and microbial activity of the polymer surface. An analysis of the structural features of the microbial communities revealed that Archaea exhibited different biodiversity at distinct temperatures. In particular, under thermophilic conditions, the relative abundance of Methanothermobacter was 56.0 %, and it plays an important role in the anaerobic degradation of PBAT/PLA/starch materials, while bacterial communities showed smaller differences. Overall, the bioplastic was able to be co-digested anaerobically with food waste to produce renewable energy. This study provides a plan for the practical application of biodegradable plastic bag collection for the combined treatment of food waste in anaerobic digesters. It provides a theoretical basis for modifications of bioplastic and domestication of anaerobic microorganisms.


Radioiodination of chicken erythrocyte histones H4 and H5 in chromatin.
The conformational state of histones in isolated chicken erythrocyte chromatin was studied using procedures developed for probing surface proteins on membranes. Under controlled conditions, only exposed tyrosyl residues react with iodide radicals, generated either by the oxidant, chloramine-T (paratoluenesulfonyl chloramide), or the enzyme lactoperoxidase, giving monoidotyrosine. Using 125-iodine, this study compared the reactive tyrosines in free and bound histones H4, and H5. The relative extent of iodination of these histones within (H4) and outside (H5) of the nucleosomes was measured after extraction and gel electrophoresis. Each of the histones was further analyzed for the extent of specific tyrosine iodination by separating the tryptic peptides by high voltage electrophoresis. The identity of the labeled peptide was determined by dansylation of the amino acids present in each hydrolyzed peptide. The results show that there is a difference in the conformational arrangement of these histones on chromatin and in the free forms, since in chromatin not all tyrosine residues are as accessible for iodination as in the denatured state. Residue 53 of histone H5 for instance is more reactive than residues 28 and 58, indicating that the segments containing the latter residues are involved in either protein-DNA or protein-protein interactions. In histone H4, preferential labeling of 2 of the 4 tyrosines present was also observed.


Anti-DNA and anti-platelet specificities of SLE-derived autoantibodies: evidence for CDR2H mutations and CDR3H motifs.
Although polyreactivity appears to be a characteristic feature of natural autoantibodies, polyreactive anti-DNA autoantibodies can be derived both from patients with autoimmune disease and from normal individuals. It is unclear whether these autoantibodies differ depending on their origin, but previous studies from our laboratory have suggested that polyreactive systemic lupus erythematosus (SLE)-derived platelet-binding anti-DNA autoantibodies have more restricted antigen reactivity and greater functional activity than normal-derived polyreactive autoantibodies. The objective of the present study was to characterize the VH and VL region sequences of 10 human hybridoma anti-DNA autoantibodies derived from peripheral blood lymphocytes of different origins [SLE, rheumatoid arthritis (RA), or normal] to determine whether there are structural differences between these autoantibodies. We show that although some unmutated germline structures (VH and VL) are represented, these are not restricted to anti-DNA autoantibodies from normal individuals and that two normal-derived anti-DNA antibodies showed quite extensively mutated VH genes. However, these mutations, unlike those found in the CDR2H of several of the SLE-derived antibodies, did not appear to be antigen-selected. Three different amino acid motifs, putatively involved in antigen binding specificity, were observed in the CDR3H segments of some of the autoantibodies. One was the previously described YYGSG motif, which was found in a normal-derived anti-DNA autoantibody, while two new potential motifs were observed only in SLE-derived platelet-binding anti-DNA autoantibodies. These data suggest that antigenic and functional differences between SLE-derived and normal-derived platelet-binding anti-DNA autoantibodies may be due to antigen-selected mutations in the CDR2H and specific amino acid motifs in the CDR3H.


Discovery of 2,5-diaminopyrimidine derivatives as the first series of selective monomeric degraders of B-lymphoid tyrosine kinase.
B-lymphoid tyrosine kinase (BLK) is an important knot of B cell receptor signaling, and regulates the function and development of B cells subset. Dysfunction of BLK is correlated with autoimmune diseases and cancer. There is an urgent need to develop selective BLK modulators to facilitate the studies of BLK in biological processes. Herein, we report the discovery of a series of 2,5-diaminopyrimidine-based compounds capable of selectively degrading BLK. The optimized compounds 9-11 possess weak biochemical inhibitory activities against BLK, yet they effectively degrade BLK and show high selectivity for BLK over other structurally and functionally related SRC family and TEC family kinases. Furthermore, compounds 9 and 11 demonstrate potent inhibitory activities in several B-lymphoid cell lines. As the first series of effective and selective monomeric BLK degraders, compounds 9-11 serve as valuable tools for further investigation of the functions of BLK.


Successful treatment of refractory recurrent vaginal candidiasis with cetirizine plus fluconazole.
To determine the prevalence of allergy in patients with recurrent vaginal candidiasis (RVC) and the efficacy of fluconazole in combination with an antihistamine drug in RVC patients who do not respond to fluconazole alone. Participants in the study included 24 women with RVC. All subjects filled out an International Study of Asthma and Allergies in Childhood questionnaire and 23 performed skin prick tests for immediate type 1 hypersensitivity as part of atopy assessment. All subjects were treated with fluconazole 150 mg weekly and were followed every 2 months for 6 months. Those who did not respond to fluconazole alone received cetirizine 10 mg daily in addition to the antifungal medication and were followed up for 6 months. The majority of women had a history of allergy (60%) and 10 of them (43%) had at least 1 positive result of a skin prick test to aeroallergens. Nineteen patients (79%) responded to fluconazole alone. The 5 patients who remained with symptoms had a medical history of allergy and obtained remission when cetirizine was associated with fluconazole. Patients with RVC refractory to standard antifungal therapy and a history of allergy may respond to the combination of antihistamine and antifungal therapy.


Adolescent sleep and technology-use rules: results from the California Health Interview Survey.
This study reports on adolescent sleep, technology-use rules at home, and their potential association using the 2017 California Health Interview Survey (CHIS). CHIS cross-sectional, representative survey data was collected between June-December 2017 via telephone. Weekday bedtimes, rise times, and night time in bed in the past week, as well as whether technology-use rules were implemented at home, were examined among 448 adolescent residents of California (mean age = 14.5, range = 12-17; 48% women). The average weekday nightly time in bed was 9.04 hours among 12- to 13-year-olds and 7.89 hours among 14- to 17-year-olds. The majority reported having technology-use rules and those who did, spent 35 more minutes in bed. Although a significant relationship was found between having technology-use rules and earlier bedtime, one was not found for time in bed, when controlling for demographics. Many California adolescents do not meet recommended sleep guidelines. Yet, technology-use rules appear to be associated with earlier bedtime. Results have implications for policy-making and adolescent sleep interventions.


Assessment of Impact of Fluoride on Mechanical Properties of NiTi and CuNiTi Orthodontic Archwires: An In Vitro Study.
The current study was carried out to assess the impact on the mechanical properties of orthodontic wires such as the nickel-titanium (NiTi) and copper-nickel-titanium (CuNiTi) wires by fluoride available in various prophylactic products. Fifty-six wire specimens were randomly divided into two groups-control group in which deionized water was used as a medium and study group in which Phos-Flur gel was used. Both study group and control group were divided into two subgroups-NiTi wire group: 0.019 × 0.025 inch NiTi archwires (14 specimens) and CuNiTi wire group: 0.019 × 0.025 inch CuNiTi archwires (14 specimens). Testing of all the wires was done under a universal force testing machine. Mean loading force among NiTi wire group and CuNiTi wire group specimens with deionized water as a medium was 682.6 and 397.4 MPa, respectively, while the mean loading force among NiTi wire group and CuNiTi wire group specimens with Phos-Flur gel as a medium was 596.1 and 368.4 MPa, respectively. While comparing between study group and control group among NiTi wires, significant results were obtained. Also, while comparing between study group and control group among CuNiTi wires, significant results were obtained. Following exposure to fluoride agents, NiTi wires and CuNiTi wires are significantly associated with reduced mechanical properties. Although fluoride acts as a vital adjunct in maintaining oral hygiene, particularly among patients undergoing fixed orthodontic treatment, its influence on the mechanical properties of the wires is an area to be explored further; thereby, its use is to be monitored.


Gas-liquid chromatographic method with electron-capture detection for the determination of residues of some phenoxyacetic acid herbicides in water as their 2,2,2-trichloroethyl esters.
A gas-liquid chromatographic method with electron-capture detection was developed for the determination of residues of 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-chloro-2-methylpenoxyacetic acid (MCPA) in water as their 2,2,2-trichloroethyl esters, and the sensitivity was compared with that for 2-chloro- and 2,2,2-trifluoroethyl esters. The residue is isolated by sorption of the phenoxy acid on Amberlite XAD-4, eluted with benzene and then esterified with 20% (v/v) 2,2,2-trichloroethanol in trifluoroacetic anhydride in the presence of sulphuric acid. The recoveries at the 0.4 ppb level are 92.1 +/- 3.4% (2,4-D) and 87.8 +/- 2.1% (MCPA) and the detection limits for a 1-1 sample of water are 0.096 ppb (2,4-D) and 0.06 ppb (MCPA).


Innovation or 'Inventions'? The conflict between latent assumptions in marine aquaculture and local fishery.
Recent European policy highlights the need to promote local fishery and aquaculture by means of innovation and joint participation in fishery management as one of the keys to achieve the sustainability of our seas. However, the implicit assumptions held by the actors in the two main groups involved - innovators (scientists, businessmen and administration managers) and local fishermen - can complicate, perhaps even render impossible, mutual understanding and co-operation. A qualitative analysis of interviews with members of both groups in the Valencian Community (Spain) reveals those latent assumptions and their impact on the respective practices. The analysis shows that the innovation narrative in which one group is based and the inventions narrative used by the other one are rooted in two dramatically different, or even antagonistic, collective worldviews. Any environmental policy that implies these groups should take into account these strong discords.


Poor Sleep Quality and Daytime Fatigue Are Associated With Subjective but Not Objective Cognitive Functioning in Clinically Relevant Hoarding.
Hoarding disorder is a chronic psychiatric condition of increasing public health concern. Recent investigation suggests a positive association between hoarding severity and insomnia symptoms. However, these findings have yet to be replicated, and the prevalence and type of sleep impairment experienced by individuals with clinically relevant hoarding symptoms (CHSs) are not known. This analysis of 20,473 members of the internet-based Brain Health Registry uses multivariate logistic regression modeling and structural equation modeling to evaluate the relationship between hoarding symptoms, sleep impairment, adverse health, and cognitive functioning. More than 12% of study participants endorsed CHSs or subclinical hoarding symptoms. After adjustment for demographic characteristics and psychiatric comorbidity, individuals with CHSs reported increased odds of sleep impairment in nearly all domains. The odds of poor sleep quality (adjusted odds ratio, 2.07; 95% CI, 1.83-2.34), sleep disturbances (adjusted odds ratio, 2.15; 95% CI, 1.91-2.43), and daytime dysfunction (adjusted odds ratio, 5.84; 95% CI, 5.12-6.65) were two- to fivefold higher for individuals with CHSs compared with those without. For all measures, the proportion of individuals reporting sleep impairment increased with hoarding severity. In our structural equation model, sleep impairment acted as a partial mediator on the indirect pathways from hoarding to subjective cognitive complaints and poorer quality of life. Identification of sleep problems among those with hoarding symptoms is a critical component of hoarding assessment. Additional research is needed to better understand the mechanisms underlying the observed relationships, including neurobiological underpinnings, and to examine the role of sleep management in treatment for hoarding behaviors.


A modification of the quininium resin test for assessing gastric acidity.
Gastric acid is an important defence against enteric infection. Studies investigating the relationship between hypochlorhydria and enteric infections or gastric malignancy have been limited by difficulties in the non-invasive measurement of gastric acidity. To develop a blood test for hypochlorhydria based on the quininium resin test. Quininium resin dissociates to liberate free quinine at pH <or= 3.5. Ten fasting adult volunteers ingested 100 mg/kg of quininium resin twice: once before (control) and once after acid suppression with rabeprazole (10 mg b.d. for 3 days). Blood and serum samples were collected at baseline and 45, 60, 75, 90 and 120 min after ingestion. Quinine levels were measured fluorometrically from 100 microL samples after simple extraction. Results obtained by the quininium resin test were compared with gastric juice pH measurements. After quininium resin ingestion, mean quinine levels were lower in the acid-suppressed than in the control condition at all times (P < 0.001). Optimized cut-off values at 90 min post-ingestion yielded 95% accuracy rates in both serum and blood. Quininium resin was well tolerated. Using blood or serum, the quininium resin test identifies hypochlorhydria accurately and non-invasively. This test may be useful in screening for gastric atrophy and in studies investigating hypochlorhydria.


Extreme thinking in clinically depressed adolescents: Results from the Treatment for Adolescents with Depression Study (TADS).
The purpose of this report is to examine relations between extreme thinking, as measured by the Dysfunctional Attitudes Scale, and the maintenance of gains among adolescents who participated in the Treatment for Adolescents with Depression Study (TADS). We examine extreme thinking among 327 adolescents (mean age=14.56, 57% female, 75% White) who received cognitive behavior therapy (CBT), fluoxetine (FLX), or a combination of CBT and FLX (COMB). Among those who met remission status on the Children's Depression Rating Scale - Revised (CDRS-R≤28; 56 at week 12, 79 at week 18) extreme thinking did not predict failure to maintain remission. This is in contrast to findings with depressed adults. Treatment influenced level of extreme thinking, and this appeared to be driven by greater endorsement of positively valenced beliefs as opposed to a decrease in negatively valenced beliefs. Developmental or investigation characteristics may account for the discrepancy in findings.


Prenatal screening for trisomy 21: a comparative performance and cost analysis of different screening strategies.
Prenatal screening for chromosome aneuploidies have constantly been evolving, especially with the introduction of cell-free fetal DNA (cfDNA) screening in the most recent years. This study compares the performance, costs and timing of test results of three cfDNA screening implementation strategies: contingent, reflex and primary. We modelled enhanced first trimester screening (eFTS) as the first-tier test in contingent or reflex strategies. cfDNA test was performed contingent on or reflex from eFTS results. A comparison was made between cfDNA screening using sequencing technology and Rolling Circle Amplification (RCA)/imaging solution. All model assumptions were based on results from previous publications or information from the Ontario prenatal screening population. At an eFTS risk cut-off of ≥1/1000, contingent and reflex cfDNA screening have the same detection rate (DR) (94%) for trisomy 21. Reflex cfDNA screening using RCA/Imaging solution provided the lowest false positive rate and cost. The number of women requiring genetic counselling and diagnostic testing was significantly reduced and women received their cfDNA screening result 9 days sooner compared with the contingent model. While primary cfDNA screening improved the trisomy 21 DR by 3-5%, it was more costly and more women required diagnostic testing. Reflex cfDNA screening is the most cost-effective prenatal screening strategy. It can improve the efficiency of prenatal aneuploidy screening by reducing the number of patient visits and providing more timely results.


A method for the evaluation of 3-hydroxyproline in the urine.
After hydrolysis of the urine by 6 M HCl, 3-hydroxyproline is purified from many interfering substances by 2 steps of chromatography on Dowex 1 X8 resin equilibrated first in the acetate form and secondly in the OH- form. The amino acid is finally evaluated by chromatography on Dowex 50 M82 resin in a Beckman multichrom amino acid analyzer. In 23 normal adult subjects the mean level was 12.5 +/- 3.5 mumol/24 h. In 8 normal children the level was 6.0 +/- 5 mumol/24 h and in 8 teenagers 15.2 +/- 2.85 mumol/24 h. The ratio of 3-hydroxyproline to 4-hydroxyproline seems indicative of a semiological value of this evaluation in cases of basement membrane collagen deterioration.


Prediction model of clinical prognosis and immunotherapy efficacy of gastric cancer based on level of expression of cuproptosis-related genes.
Gastric cancer is one of the most common malignancies in the world and ranks fourth among cancer-related causes of death. Gastric adenocarcinoma is the most common pathological type of gastric cancer; usually, this tumor is associated with distant metastasis upon first diagnosis and has a poor prognosis. Cuproptosis is a novel mechanism of cell death induced by copper, and is closely related to tumor progression, prognosis and immune response. However, the role of cuproptosis-related genes (CRGs) in the tumor microenvironment (TME) of gastric cancer has yet to be elucidated. Gastric adenocarcinoma data were downloaded from the Cancer Genome Atlas (TCGA) database. Through bioinformatics analysis, a risk scoring model was constructed from cuproptosis gene-related lncRNA. Then, we investigated the relationship between prognosis and the TIME of gastric cancer according to clinical characteristics and risk score. Validation of the model showed that the overall survival (OS) of the high-risk group was significantly lower than that of the low-risk group (P < 0.001) and that the risk score was an independent predictor of prognosis (P < 0.001). The new model was significantly correlated with the prognosis and TIME of patients with gastric cancer, including immune cell infiltration, tumor mutation burden (TMB) score, targeted drug sensitivity, and immune checkpoint gene expression. In addition, a prognostic nomogram was established based on the risk score (AC008915.2, AC011005.4, AC023511.1, AC139792.1, AL355312.2, LINC01094 and LINC02476). Our analysis revealed that the prognostic model of cuproptosis-related genes could effectively predict the prognosis of patients with gastric cancer and comprehensively establish the relationship between cuproptosis genes and tumor immunity. This may provide a new strategy for the precise treatment of gastric cancer.


Nonylphenol-induced Ca2+ elevation and Ca2+-independent cell death in human osteosarcoma cells.
The effect of the environmental toxicant nonylphenol on cytosolic free Ca2+ concentration ([Ca2+]i) and proliferation has not been explored in human osteoblast-like cells. This study examined whether nonylphenol alters Ca2+ levels and causes cell death in MG63 human osteosarcoma cells. [Ca2+]i and cell death were measured using the fluorescent dyes fura-2 and WST-1 respectively. Nonylphenol at concentrations above 3 microM increased [Ca2+]i in a concentration-dependent manner. The Ca2+ signal was reduced by 90% by removing extracellular Ca2+. The nonylphenol-induced Ca2+ influx was insensitive to blockade of L-type Ca2+ channel blockers. After pretreatment with 10 microM nonylphenol, 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) failed to induce [Ca2+]i rises. Inhibition of phospholipase C with 2 microM U73122 did not change nonylphenol-induced [Ca2+]i rises. The nonylphenol-induced [Ca2+]i rises were enhanced or inhibited by phorbol myristate acetate or GF 109203X, respectively. At concentrations of 10 and 20 microM nonylphenol killed 55% and 100% cells, respectively. The cytotoxic effect of 10 microM nonylphenol was unaltered by pre-chelating cytosolic Ca2+ with BAPTA. Collectively, in MG63 cells, nonylphenol induced [Ca2+]i rises by causing Ca2+ release from intracellular stores and Ca2+ influx from extracellular space. Furthermore, nonylphenol can cause Ca2+-unrelated cytotoxicity in a concentration-dependent manner.


The RBE of euoxic and hypoxic Ehrlich ascites tumour cells irradiated with negative pions in vitro and in vivo.
Ehrlich ascites tumour cells (EAT-F5) were irradiated in vitro with the sparsely ionizing "plateau pions" and with the more densely ionizing "peak pions". From cell survival curves obtained under euoxic and hypoxic conditions, the RBE for the production of irreparable lethal and potentially lethal damage was derived. Cells of the same strain were used to produce solid tumours in the leg muscle of mice. Survival of these cells irradiated in vivo was compared with the corresponding in vitro data. It can be concluded that a large fraction of hypoxic cells is present in these tumours and that repair takes place in vivo at least as much as under the in vitro conditions. These solid tumours were irradiated with 5 dose fractions of fast protons as reference radiation, and the probability pT of tumour destruction was determined as a function of dose. Irradiation with a spot of "plateau pions" indicated the same pT as a function of dose, but side effects in the intestinal tract increased also with dose, leading to the death of the animals. The probability for avoiding these side effects in the animals, pS, was also determined. Using both values, pT and pS, the probability for local tumour control was estimated. Irradiation with "peak pions" resulted in an RBE of about 2.5 for tumour destruction corresponding to the RBE found in vitro for the production of irreparable lethal lesions in hypoxic cells. Side effects in the mouse gut showed a RBE of 2.0, corresponding to irreparable lethal lesions in euoxic in vitro cells.


The Werther effect following the suicides of three korean celebrities (2017-2018): an ecological time-series study.
The suicide rate in Korea was the highest among the member countries of the Organization for Economic Cooperation and Development(OECD) for 2013-2016 and 2018-2020. In korea, suicide was the leading cause of death among individuals aged 10-39, and the second leading cause of death for aged 40-59. Thus, this study aimed to examine the Werther effect of the suicides of three Korean idol singers (Jonghyun: December 18, 2017, Sulli: October 14, 2019, and Hara Gu: November 24, 2019). The study conducted Poisson regression and used the cause-of-death statistics microdata from 2016 to 2020 provided by Statistics Korea. The case periods ranged from the day of the suicide of each celebrity to 10 weeks after. The control periods were all weeks from 2016 to 2020, excluding the case periods. The suicide rates in Korea significantly increased by 1.21, 1.30, and 1.28 times after the deaths of Jonghyun, Sulli, and Hara Gu, respectively. The Werther effect was more evident in women than men. Suicide rate among individuals aged 10-29 years was greater than those for other age groups. This study confirmed that the rate of copycat suicides increased after three celebrity singers in Korea died by suicide. Nevertheless, the rate of suicide after the suicide of the three celebrity singers was lower than those in previous studies in Korea.


Late outcomes of pediatric heart transplantation are independent of pre-transplant diagnosis and prior cardiac surgical intervention.
An increasing number of children are being referred for cardiac transplantation after (1) failing conventional corrective or palliative surgical reconstruction, (2) after stabilization with mechanical circulatory support devices, and (3) when primary graft failure or advanced cardiac allograft vasculopathy are established. The records of 417 infants and children (age range, 0-18 years) who underwent cardiac transplantation from November 1985 through December 2005 at Loma Linda University Children's Hospital were retrospectively reviewed. The pre-transplantation diagnosis was used to divide patients into 3 groups: primary cardiomyopathy (CM), 103; hypoplastic left heart syndrome (HLHS), 154; and other complex congenital heart disease (CCHD), 160. These groups were compared and analyzed for differences in early and late morbidity and mortality. Operative mortality was significantly lower in the CM group compared with the HLHS (p < 0.02;) and CCHD groups (p < 0.01). Long-term actuarial recipient survival, however, was similar for all groups. The 15-year actuarial survival was 59% for the CM Group, 57% for the HLHS Group, and 50% for the CCHD Group. Actuarial survival after retransplantation is not statistically different from that with primary cardiac transplantation. Although peri-operative survival was lower in infants and children with HLHS and CCHD compared with those with CM, long-term survival has been the same for all groups. Late survival after retransplantation was not statistically different than among those with primary cardiac transplantation.


Normative size of the osseous part of calcaneal bursa and its comparison with other calcaneal articular areas.
The retro-calcaneal bursa presents a synovial and a non-synovial osseous part of variable dimensions. Studies objectively measuring the variability of the size of this osseous bursal surface cannot be found in literature. The objective of this study was to investigate (i) the dimension variability of the bony part of the bursa and (ii) the relationship of this surface to other articulating areas of the calcaneus. A digital planimeter was used to measure the bursae (n=86) and other articular surface areas of the calcaneus and statistically compared with ANOVA and correlation estimations. The osseous area measured 1.12 (±0.55) cm2, with only the superior articulating area demonstrating a weak correlation to this osseous surface. The osseous area presents a weak correlation with the axial articulating area of the calcaneus. Information on the size of the bony bursa may help safe excision of retrocalcaneal exostoses and in Achille's tendon repair around the posterior tuberosity.


Impact of agricultural farms on the environment of the Puck Commune: Integrated agriculture calculator-CalcGosPuck.
Leaching of nutrients from agricultural areas is the main cause of water pollution and eutrophication of the Baltic Sea. A variety of remedial actions to reduce nitrogen and phosphorus losses from agricultural holdings and cultivated fields have been taken in the past. However, knowledge about the risk of nutrient leaching has not yet reached many farmers operating in the water catchment area of the Baltic Sea. The nutrient balance method known as "At the farm gate" involves calculating separate balances for nitrogen (N), phosphorus (P) and potassium (K). After estimating all the components of the nutrient balance, the total balance for NPK is calculated and the data obtained is expressed as the ratio of total change (surplus) to the area of arable land on a farm. In addition, the nutrient usage efficiency on a farm is also calculated. An opinion poll was conducted in 2017 on 3.6% (n = 31) of the farms located in commune of Puck. The total area of the farms including arable and grass land ranged from 5 to 130 ha with an average of 45.82 ha. The arable land was on average 30.79 ha ranging from 4.45 to 130 ha while the grassland averaged 12.77 ha and ranged from 0 to 53 ha. The average consumption of mineral fertilizer in the sample population of farms was 114.9 kg N, 9.3 kg P, and 22.9 kg K·ha-1of agricultural land (AL), respectively. N balance in the sample farms being ranged from -23.3 to 254.5 kg N·ha-1AL while nutrient use efficiency ranged from 0.40% to 231.3%. In comparison, P surplus in the sample farms was 5.0 kg P·ha-1AL with the P use efficiency of 0.4-266.5%. Mean N fertilizer consumption in the tested farms was higher than the average usage across Poland and in the Pomeranian Voivodeship. However, mean consumption of potassium fertilizers was lower than mentioned averages. Mean P fertilizer consumption was higher than in the Pomeranian Voivodeship, but lower compared to the entire country. Generally, on the basis of designated research indicators of farm pressures on water quality, concentrations of total nitrogen and total phosphorus were obtained. CalcGosPuck (an integrated agriculture calculator) will help to raise farmers' awareness about NPK flow on farm scale and to improve nutrient management.


Maintenance of Golgi apparatus structure in the face of continuous protein recycling to the endoplasmic reticulum: making ends meet.
I focus here on the Golgi apparatus and the dynamic relationship between the Golgi apparatus, the central organelle of the secretory pathway, and the endoplasmic reticulum (ER). The proteins and lipids of the Golgi apparatus originate in the ER, and cargo proteins and lipids that also originate in the ER are processed and sorted within the Golgi apparatus. The Golgi apparatus is indeed the central organelle of the secretory pathway. Surprisingly, many, if not all, of the proteins and accompanying lipids of the Golgi apparatus cycle continuously between the Golgi and the ER. Neither the Cisternal Maturation nor the Vesicular Transport/Stable Compartment model of Golgi apparatus function predicts continuous cycling of Golgi resident proteins through the ER. Evidence for this cycling comes from multiple experimental approaches, including ER-exit block-revealed accumulation of recycled Golgi resident proteins in the ER, evidence for exchange of green fluorescent protein (GFP)-tagged Golgi proteins or their analogues between Golgi and ER pools, and cisternal rab overexpression-induced redistribution of Golgi resident proteins to the ER. The implications of Golgi protein cycling for the maintenance of Golgi structure in the interphase mammalian cell are discussed. The challenge for the future is to put Golgi resident protein cycling pathway(s) to protein machinery and to characterize the cumulative, weak, dynamic interactions that hold the Golgi apparatus together. In doing so, new paradigms of organelle biogenesis will emerge.


"Double-orifice" technique to repair extensive mitral valve excision following acute endocarditis.
The use of conservative surgical techniques to treat mitral valve regurgitation secondary to acute endocarditis is controversial. Reconstruction of the anterior leaflet may represent an additional challenge in such a setting. We report a case of mitral valve repair where extensive excision of the anterior leaflet and related chordae tendinea was necessary because of large vegetation secondary to acute endocarditis. The "double-orifice" technique was performed and allowed the salvage of the native valve. There was no recurrent infection at 6 months from surgery, with optimal hemodynamic results.


The origin of the initial heat associated with a single impulse in mammalian non-myelinated nerve fibres.
1. A study has been made of the temperature changes associated with the passage of a single impulse in rabbit desheated vagus nerves.2. The initial changes consist of an evolution of positive heat followed by a reabsorption of most of it; i.e. there is a phase of positive and a phase of negative heat production.3. The size of the positive heat, its time of onset, and the time of onset of the negative heat have been measured by an analogue method of analysis. In addition, these parameters, together with the size of the negative heat and the duration of both phases of initial heat, have been studied with the aid of a computer, and also by conventional heat block analysis.4. At about 5 degrees C the measured positive heat is 7.2 mucal/g. impulse. It starts as soon as the compound action potential reaches the thermopile and lasts for about 107 msec.5. This positive heat decreases with increasing temperature, the ratio of heat at 4 degrees C to that at 14 degrees C being 1.86.6. The measured negative heat at about 5 degrees C is 4.9 mucal/g. impulse. It starts 102 msec after the onset of positive heat, and lasts for about 240 msec.7. When the sodium of Locke solution is replaced by lithium the positive heat is reduced by 19%, but the negative heat is increased by 22%.8. In potassium-free solutions the positive heat is hardly affected (increase of 5%), but the negative heat is more than doubled. As a result the nerve may become briefly colder than its initial temperature by about 2 mu degrees C.9. The effect of sodium-deficient solutions on the positive heat is somewhat variable, but the negative heat is consistently diminished.10. Replacement of the chloride of Locke solution by sulphate or nitrate has little effect on the positive heat. The negative heat is reduced in size by 26% and in duration by 22%.11. Replacement of most of the sodium of Locke solution by barium reduces or abolishes the negative heat, and increases the measured size of the positive heat nearly threefold.12. Veratrine (10(-5) g/ml.) produces a nearly tenfold increase in the net positive heat.13. Ouabain (1 mM) and antimycin A (1 mug/ml.) applied for 30-60 min have little effect on the initial heat production.14. Over the temperature range 5-15 degrees C, and for the ionic solution changes described above, there is close agreement in timing between the positive heat and the rising phase of the action potential, and between the negative heat and the falling phase.15. Because of the inevitable temporal dispersion of the action potential over the face of the thermopile, the observed temperature changes are smaller than those which occur at a single point in the nerve close to a stimulating electrode. The corrected value of the positive heat at 5 degrees C is 24.5 mucal/g. impulse, while that of the negative heat is 22.2 mucal/g. impulse.16. The heats of mixing of the ions in solution that interchange during the action potential are much too small to account for the observed initial heats, but an exchange of ions associated with fixed charges might make a significant contribution to the heats.17. The condenser theory, according to which the positive heat represents the dissipation of electrical energy stored in the membrane capacity, while the negative heat results from the recharging of the capacity, appears unable to account for more than half of the observed temperature changes.18. It seems probable that the greater part of the initial heat results from changes in the entropy of the nerve membrane when it is depolarized and repolarized.


Value of tissue Doppler echocardiography in children with pulmonary hypertension.
The impact of pulmonary hypertension (PHT) on right ventricular and left ventricular (LV) function in children with PHT is unknown, and echocardiographic data combining conventional and Doppler tissue imaging (DTI) on PHT in children are sparse. Forty-one children (18 male; mean age, 7.9 ± 5.6 years) with PHT and structurally normal hearts (27 with idiopathic PHT, 14 with associated PHT) and 44 age-matched healthy controls were assessed using conventional echocardiography and DTI. Children with PHT had enlarged tricuspid valve diameters, right atrial areas, pulmonary artery dimensions, and LV eccentricity indices. In addition, pulmonary acceleration time and tricuspid annular plane systolic excursion were significantly reduced in patients compared with controls. DTI revealed that children with PHT had significantly lower systolic (S) and early diastolic (E) velocities at the tricuspid and septal levels. Despite preserved LV ejection fractions, left lateral free wall systolic velocities were significantly reduced in patients with PHT. Significantly reduced LV rapid filling velocities (E) suggested an underloaded left ventricle or LV diastolic dysfunction in children with PHT compared with controls. Pulmonary acceleration time and tricuspid annular plane systolic excursion correlated best with DTI systolic tricuspid and septal velocities. Despite not being evident on conventional two-dimensional echocardiography, LV systolic performance appears to be impaired in children with PHT. Quantitative DTI assessment of ventricular function and ventricular-ventricular interactions in this setting might provide further insights into the mechanisms leading to end-stage PHT and may guide clinicians to optimize antifailure treatment.


Amelioratory effects of testosterone treatment on cognitive performance deficits induced by soluble Aβ1-42 oligomers injected into the hippocampus.
This study was undertaken to investigate the protective effects and potential mechanism of testosterone (T) on cognitive performance in adult male rats given bilateral intrahippocampal injections of beta amyloid 1-42 oligomers (Aβ1-42) combined with gonadectomy (Aβ+GDX). A series of experiments were designed to verify the optimal administration time and dose of T and to explore its potential protective mechanisms on spatial ability in Aβ+GDX rats in the Morris water maze test. Aβ1-42 was injected only once two weeks before testing, while T and the androgen receptor (AR) antagonist flutamide (F) were administered daily beginning 2 days before and throughout the 6 days of testing. The Aβ1-42 injection and GDX individually impaired cognitive performance, and the combination of these treatments was additive, leading to even greater impairment. The serum T level peaked at 48 h after administration. T doses ranging from 0.25 to 1.00 mg corresponding to serum T levels of 4.5-21.35 ng/ml improved the spatial ability. Animals administered 0.75 mg of T corresponding to the serum T level of 15.2 ng/ml had the most significantly improved behavioral performances. However, higher T doses of 1.50 and 2.00 mg resulting in serum T levels of 34.8 and 45 ng/ml, respectively, impaired the behavioral performances. F had no effect on the serum T level and spatial ability, but it blocked the activational effect of T. These findings indicate that the effect of T on behavioral performances is partly mediated through ARs.


Patients' expectations for follow-up in breast cancer--a preliminary, questionnaire-based study.
The current National Institute for Clinical Excellence (NICE) guidelines recommends that only 2-3 years of follow-up after breast cancer be provided. Clinicians are unwilling to implement these guidelines. However, little has been done to establish patients expectations before they embark on their regular follow-up programme. We have sought the opinions of patients in a questionnaire-based prospective cohort study. Expectations for length and frequency of follow-up were established in women prior to attending their first scheduled follow-up review 1 year after treatment. In addition, patients were asked their opinions on what clinics were designed to achieve. An attempt was made to establish whether patients would be happy with less follow-up when informed of the inefficiency of routine clinic visits. Most women expect some follow-up, but expectations for length and frequency vary dramatically. Most believe follow-up is for the detection of relapse, but very few see psychological support or side effect detection as being central to clinicians' aims. One third of women would be happy to not come back to clinic at all when told how infrequently routine clinical examination detects metastatic disease.


Excessive variability in systolic blood pressure that is self-measured at home exacerbates the progression of brain white matter lesions and cognitive impairment in the oldest old.
To investigate the effects of variability in self-measured systolic blood pressure at home on the progression of cognitive impairment and white matter lesions in the oldest old. Between April 2009 and October 2009, 248 oldest old aged 80 years or older were eligibly enrolled from geriatric practices and community-dwelling areas of Shandong, China. Self-measured blood pressure at home (HBP) was measured for 7 consecutive days at the baseline, and the Mini-Mental State Examination (MMSE) score and brain white matter hyperintensities (WMH) were assessed at the baseline and during the final follow-up visit. Variability in systolic HBP was evaluated using coefficient of variation (CV) in serial daily systolic HBP measurements of the last 6 consecutive days. After an average of 2.3 years of follow-up visits, 232 oldest old were included in and 16 were excluded from the analysis. The MMSE score declined -4.76 (interquartile ranges: -10.71, -0.83) %, the periventricular WMH, deep WMH, total WMH and WMH fraction increased 16.46 (s.d.: 6.72)%, 10.05 (s.d.: 6.40)%, 14.69 (s.d.: 6.07)% and 15.95 (s.d.: 6.32)%, respectively, in the total oldest old. A declined percentage of the MMSE score and increased percentages of the periventricular WMH, deep WMH, total WMH and WMH fraction in the high group divided by tertile of the CV of the systolic HBP at baseline were greater than those in the low group (P<0.05). The significant differences were retained after adjusting for covariates, including the MMSE score, periventricular WMH, deep WMH and WMH fraction at the baseline (P<0.05). Excessive variability in self-measured systolic HBP exacerbates the progression of cognitive impairment and brain white matter lesions in the oldest old.


Cochlear implantation in prelingually deafened children with residual hearing.
The purpose of this study was to examine the speech perception skills of prelingually deafened French children with preoperative residual hearing who received multichannel cochlear implants. The design of the study incorporated a within-subject, repeated measures design for assessing speech perception skills. Montpellier, Toulouse and Lyon Pediatric Cochlear Implant Centers. Seven prelingually deafened children demonstrating marginal benefit from conventional amplification prior to implantation with a Nucleus multichannel cochlear implant, served as subjects for the speech perception assessment (a speech recognition score less than 30% defines marginal benefit from acoustic amplification on open set materials). The mean age at implantation was 7 years, 9 months. Speech perception skills were assessed using open set materials and the MUSS and MAIS questionnaires. Open-set speech recognition averaged 21.4% before implantation, and 83.6% after 1 year's cochlear implant experience. All children demonstrated an open-set score over 60% after 12 months of CI use. MAIS test scores averaged 18.1/40 before implantation and 35.1/40 after 9 months of CI use. MUSS test scores averaged 24.4/40 before implantation and 34.1/40 after 9 months of CI use. Cochlear implantation should be considered for prelingually hearing impaired children demonstrating marginal benefit from hearing aids, with a speech recognition score less than 30% on open set materials, in order to improve their speech discrimination skills.


Removal of 2,4-dinitrophenol using hybrid methods based on ultrasound at an operating capacity of 7 L.
Sonochemical removal of 2,4-dinitrophenol (DNP) has been investigated using ultrasonic bath, with an operating capacity of 7 L, fitted with a large transducer with longitudinal vibrations having a 1 kW rated power output and operating frequency of 25 kHz. It has been revealed from calorimetric studies that maximum power is dissipated at a capacity of 7 L. The concentration of DNP has been monitored with an objective of evaluation of the efficacy of ultrasonic reactor in combination with process intensifying approaches for the removal of DNP. The effect of operating pH and additives such as hydrogen peroxide and ferrous iron activated persulfate on the extent of removal of DNP has been investigated. It has been observed that the extent of removal is greater at lower pH (pH 2.5 and 4) than at higher pH (pH 10). The combined treatment strategies such as ultrasound (US)/Fenton, US/advanced Fenton and US/CuO/H2O2 have also been investigated with an objective of obtaining complete removal of DNP using hybrid treatment strategies. The extent of removal has been found to increase significantly in US/Fenton process (98.7%) as compared to that using US alone (5.8%) which demonstrates the efficacy of the combined process. First order kinetics has been fitted for all the approaches investigated in the work. Calculations of cavitational yield indicated the superiority of the reactor design as compared to the conventional ultrasonic horn type reactors. The main intermediates formed during the process of removal of DNP have been identified.


Clinical and microbiological outcomes in treatment of men with non-gonococcal urethritis with a 100-mg twice-daily dose regimen of sitafloxacin.
Several microorganisms cause non-gonococcal urethritis (NGU). Failure to eradicate Mycoplasma genitalium from the urethra could be associated with persistent or recurrent urethritis; thus, the choice of antibiotics with activities potent enough to eradicate M. genitalium is crucial in the treatment of NGU. In in vitro studies, sitafloxacin has been shown to be highly active against Chlamydia trachomatis and M. genitalium. We treated 89 males with NGU, including 15 patients with persistent or recurrent NGU and 1 patient with post-gonococcal urethritis, with a 100-mg twice-daily dose regimen of sitafloxacin to assess its efficacy against NGU. We examined first-void urine samples for the presence of C. trachomatis, M. genitalium, Ureaplasma parvum, and Ureaplasma urealyticum. After treatment, we evaluated 73 patients for clinical outcomes and 44 for microbiological outcomes. Symptoms were alleviated in 62 (84.9%) patients, who were judged clinically cured. Microorganisms detected before treatment were eradicated in 42 (95.5%) patients, who were judged microbiologically cured. Regarding microbiological outcomes of specific microorganisms, eradication rates of C. trachomatis (n = 33), M. genitalium (n = 11), and U. urealyticum (n = 10) were 100%, 100%, and 80.0%, respectively. In all 5 patients with M. genitalium-positive persistent or recurrent NGU who had experienced treatment failures with antibiotics, the mycoplasma was eradicated. These results suggested that the sitafloxacin regimen used, which was effective on both M. genitalium and C. trachomatis infections, could be useful as an appropriate option as first- and second-line treatment of NGU.


Intracisternal injection of CRF antagonist blocks surgical stress-induced inhibition of gastric secretion in the rat.
The effects of intracisternal injection of CRF antagonist, alpha-CRF 9-41, on the inhibition of gastric acid secretion elicited by intracisternal injection of corticotropin-releasing factor (CRF) and stress were investigated in conscious pylorus-ligated rats. Intracisternal injection of the alpha-helical CRF 9-41 (50 micrograms) did not influence basal gastric secretion, but injected concomitantly with intracisternal CRF (5 micrograms), completely blocked CRF (5 micrograms)-induced inhibition of gastric secretory volume, acid concentration and output. Intracisternal injection of alpha-helical CRF 9-41 (3, 10, 50 micrograms) produced a dose-related reversal (0, 52 and 100%) of brain surgery-induced inhibition of gastric acid output. By contrast intravenous injection of CRF antagonist (50 micrograms) did not inhibit gastric hyposecretory response to brain surgery. These data suggest that endogenous CRF in the brain may mediate stress-induced gastric hyposecretion in the rat.


Systemic deficiency of mouse arachidonate 15-lipoxygenase induces defective erythropoiesis and transgenic expression of the human enzyme rescues this phenotype.
Arachidonic acid 15-lipoxygenases (ALOX15) are lipid peroxidizing enzymes, which has previously been implicated in the maturational breakdown of intracellular organelles and plasma membrane remodeling during reticulocyte-erythrocyte transition. Conventional Alox15-/- mice are viable, develop normally but do not exhibit a major defective erythropoietic phenotype. To characterize the putative in vivo relevance of Alox15 for red blood cell development, we explored the impact of systemic inactivation of the Alox15 gene on mouse erythropoiesis. We found that Alox15-/- mice exhibited reduced erythrocyte counts, elevated reticulocyte counts and red cell hyperchromia. The structure of the plasma membrane of Alox15-/- erythrocytes is altered and a significant share of the red cells was present as echinocytes and/or acanthocytes. An increased share of the Alox15-/- erythrocytes cells were annexin V positive, which indicates a loss of plasma membrane asymmetry. Erythrocytes of Alox15-/- mice were more susceptible to osmotic hemolysis and exhibited a reduced ex vivo life span. When we transgenically expressed human ALOX15 in Alox15-/- mice under the control of the aP2 promoter the defective erythropoietic system was rescued and the impaired osmotic resistance was normalized. Together these data suggest the involvement Alox15 in the maturational remodeling of the plasma membrane during red cell development.


Clinical Characteristics and Prognostic Significance of Subclinical Seizures in Focal Epilepsy: A Retrospective Study.
The aim was to evaluate the clinical characteristics and prognostic significance of subclinical seizures (SCSs) on scalp video-electroencephalogram (VEEG) monitoring with or without intracranial electroencephalogram (IEEG) monitoring in patients who had epilepsy surgery. We reviewed 286 epileptic patients who underwent subsequent epilepsy surgery during scalp-VEEG evaluation with or without IEEG monitoring between 2013 and 2020, with a minimum follow-up of 1 year. The prevalence and clinical characteristics of SCSs, as well as their prognostic significance, were analyzed. A total of 286 patients were enrolled for analysis, and 80 patients had IEEG implanted. SCSs were recorded in 9.79% of the patients based on VEEG and 50% based on IEEG. In the VEEG group (n = 286), younger seizure onset (P = 0.004) was associated with the presence of s-SCSs (SCSs detected on scalp VEEG). In the IEEG group (n = 80), temporal lobe epilepsy (P = 0.015) was associated with the presence of i-SCSs (SCSs detected on IEEG). Of 286 patients, 208 (72.73%) were seizure-free in the VEEG group, and 56 0f 80 patients (70%) were seizure-free in the IEEG group through the last follow-up. In the VEEG group, the presence of s-SCSs did not affect seizure outcome; predictors of seizure recurrence were longer epilepsy duration (P = 0.003, OR 1.003, 95% CI 1.001-1.005), history of focal to bilateral tonic-clonic seizure (P = 0.027, OR 1.665, 95% CI 1.060-2.613), nonspecific pathology (P = 0.018, OR 2.184, 95% CI 1.145-4.163), and incomplete resection (P = 0.004, OR 2.705, 95% CI 1.372-5.332). In the IEEG group, i-SCSs were significantly associated with seizure outcome (P = 0.028, OR 0.371, 95% CI 0.153-0.898). The rate of SCSs captured on IEEG monitoring was higher than that on VEEG monitoring during presurgical evaluation. SCSs detected on VEEG monitoring were associated with younger seizure onset. SCSs detected on IEEG monitoring were associated with temporal lobe epilepsy and also predicted surgical outcomes in focal epilepsy.


Immunocytochemistry and heterogeneity of rat brain vimentin.
In unfixed cryostat sections of the brains of early postnatal and adult rats, we screened for cells containing vimentin-positive intermediate filaments (VI+-IFs) by applying a panel of four monoclonal antibodies (Mabs VI-01, VI-02, VI-05 and VI-5B3) using indirect immunofluorescence. All of the Mabs stained VI+-IFs in the stromal part of the choroid plexus, in endothelial cells of blood vessels and in meninges in both adult and immature brains, although with varying strength (VI-5B3 and VI-01 stained more strongly than VI-05 and VI-02). In the brain parenchyma of adults, intense staining was mainly localized in ventricular ependymal cells (VI-5B3/VI-01 greater than VI-02/VI-05) and fibrous astrocyte-like cells (FAs). In the immature brain, the ependymal cells were activated in appearance, with evidence of cell enlargement, greater spreading of VI+-IFs within the cytoplasm and more pronounced VI+ cytoplasmic protrusions into the brain parenchyma. VI+-FAs were found near the ependymal and meningeal borders as well as in the white matter tracts of adult brain (VI-5B3/VI-01 greater than VI-05 greater than VI-02). In immature animals, VI+-FAs were less frequently encountered in the forebrain regions, except in and near the subepenydmal layer (in the adjacent parenchyma) as well as in submeningeal layers. Weaker staining was usually elicited by Mabs VI-02 and VI-05. In the cerebellum, Bergmann cell fibers were stained in both age groups. In adults, the most intense fluorescence usually occurred in segments close to the pia (VI-5B3/VI-01 greater than VI-05 greater than VI-02).(ABSTRACT TRUNCATED AT 250 WORDS)


It's time for a rethink.
Recently, I heard about a child who was referred to specialist obesity services by the school nurse, despite being considered by the family to be fit, healthy, happy and a normal size.


Effects of small mammal burrows on water infiltration in a cool desert environment.
The hypothesis that small mammal burrows can increase the amount of water infiltrating into the soil profile was tested. The amount of water added to the soil profile from spring recharge in areas adjacent to ground squirrel (Spermophilus townsendii and S. elegans) burrows was compared to nearby areas without burrows. Recharge amounts in burrow areas were significantly higher than nonburrow areas. An average of 21% more of the winter precipitation infiltrated into the soil near burrows. The amount of recharge was also found to be positively related to burrow density. Burrows also affected the distribution of the recharge by adding significantly more water to the deeper portions (>50 cm) of the soil profile.


Centchroman altered the expressions of tumor-related genes through active chromatin modifications in mammary cancer.
Centchroman (CC), a female oral contraceptive, has been shown to possess breast anti-cancer activities. Recently, we have shown CC-mediated antimetastatic effect through reversal of epithelial-to-mesenchymal transition (EMT) in breast cancer. The loss of tumor suppressor genes (TSGs) has been shown to promote EMT in breast cancer. Therefore, in the present study, we investigated the effect of CC-treatment on the expression of tumor-related genes including both tumor suppressor- and tumor promoter genes in breast cancer. CC treatment resulted in G0 /G1 phase cell cycle arrest in human breast cancer MDA-MB-231, SK-BR-3, and ZR-75-1 cells with the concomitant induction of TSGs such as p21WAF1/CIP1 , p16INK4a , and p27Kip1 . In addition, CC treatment also resulted in the downregulation of tumor promoter gene, human telomerase reverse transcriptase (hTERT). The induction of TSGs and downregulation of hTERT was found to be correlated with decreased expression levels of histone deacetylases (HDACs) and DNA methyltransferases (DNMTs). Further, mechanistic studies revealed CC-induced global DNA demethylation and alterations in the enrichment of chromatin modification markers at the promoters of p21 and hTERT. These in vitro results were corroborated with in vivo findings in 4T1-syngeneic mouse model, where CC-treatment resulted in tumor growth reduction accompanied with the induction of TSGs and alterations in the expression levels of HDACs, DNMT1, and histone modification markers. Overall, our findings suggest that CC-treatment induces the expression of TSGs and downregulates hTERT through histone modifications and DNA methylation changes. Therefore, CC could be further developed into a promising drug candidate against breast cancer. © 2015 Wiley Periodicals, Inc.


Heparin improves alveolarization and vascular development in hyperoxia-induced bronchopulmonary dysplasia by inhibiting neutrophil extracellular traps.
The objective of this study was to assess the role of NETs in BPD of hyperoxia-induced rat model and the effect of heparin on alveolarization and vascular development in BPD. The neonatal rats exposed to 90% oxygen continuously for 7 days to mimic BPD, meanwhile, the rats were injected by different doses of histones to evaluate the impact on lung injury. The newborn rats exposed to hyperoxia were injected by different doses of heparin (250 U/kg, 500 U/kg) or anti-H4 antibody to evaluate the effect of heparin. Histones and hyperoxia impaired alveolarization with the increase of mean linear intercept (MLI) and the decrease of radial alveolar count (RAC), decreased lung angiogenesis with the decrease expression of VEGF, and increased the expression of NETs, histones and pro-inflammatory factor. However, low dose heparin (250U/kg) administration enhanced survival, improved alveolarization and vascular development in hyperoxia-induced BPD, as well as reduced expression of NETs, histones and pro-inflammatory factor. We concluded that heparin improves alveolarization and vascularization in BPD by inhibiting NETs.


The role of vitamin E in T-cell differentiation and the decrease of cellular immunity with aging.
Spontaneously hypertensive rats (SHR) as a model for aging were used in this experiment and fed a regular (50 IU/Kg diet) or high vitamin E (500 IU/Kg diet) diet for 6 weeks. At 12 weeks old, they were killed and assayed. Although proliferation of thymic lymphocytes was significantly decreased in SHR fed the regular diet compared to Wistar Kyoto rats (WKY) fed the same diet, high vitamin E diet enhanced proliferation of thymic lymphocytes in SHR to almost the levels in WKY fed the regular diet. In addition, the expressions of both CD4 and CD8 antigens on CD+CD8+ T cells, immature T cells existing in thymic cortex, were also decreased in SHR, and significantly improved by high vitamin E diet. These results suggest that high vitamin E diet enhances thymic lymphocyte proliferation through increased T-cell differentiation in thymus. Then, the effect of vitamin E on T-cell differentiation in thymus was investigated by using male Fisher rats. Rats were divided into three groups; vitamin E-free, regular and high vitamin E groups and fed a diet containing various levels of vitamin E (0, 50 and 500 IU/Kg diet) for 7 weeks. Although the percentages of CD4+CD8- and CD4-CD8+ T cells in thymocytes were significantly greater in the high vitamin E group, the percentage of CD4+CD8- T cells inversely decreased in the vitamin E-free group compared to the regular group. We have tried to investigate the mechanism of the increased T-cell differentiation in thymus of rats fed the high vitamin E diet through cytokine production, and thymic epithelial cell (TEC) and macrophage functions. We have found that vitamin E enhances T-cell differentiation through the increase of not macrophage but TEC function in thymus, which is associated with the increased binding capacity of TEC to immature T cells via increased expression of adhesion molecule, ICAM-1. These results suggest that vitamin E is a potent nutrient for promoting health in the aged via the improvement of cellular immunity decreased with aging.