Nicotine binds to the transthyretin-thyroxine complex and reduces its uptake by placental trophoblasts.
A supply of maternal thyroid hormone (thyroxine, T4) is essential for normal human fetal development. Human placental trophoblasts synthesize, secrete and take up the T4 binding protein transthyretin, providing a route for maternal T4 to enter the placenta. Transthyretin is also involved in T4 transport in other tissues such as the brain choroid plexus. Nicotine alters transthyretin synthesis and function in rat choroid plexus. If nicotine influences trophoblast turnover of transthyretin, then it may directly affect placental transfer of T4 to the developing fetus and contribute to the negative impacts of smoking on fetal growth, development and placental function. The effect of nicotine on trophoblast uptake of Alexa-labelled transthyretin was measured using live cell imaging. The effect of nicotine on protein expression was measured by western blotting. Interactions between transthyretin, T4 and nicotine were investigated using chemical cross-linking techniques and molecular dynamic simulations. Nicotine blocks uptake of transthyretin-T4 by human placental trophoblast cells. Nicotine reduces the expression of the trophoblast scavenger receptor class B type 1 (SR-B1) that plays a role in transthyretin-T4 uptake. Molecular dynamic modelling suggests that when T4 is bound to transthyretin, nicotine binding increases tetramer stability, reducing the ability of the transthyretin-T4 complex to enter trophoblast cells. Our data suggest that nicotine exposure during pregnancy reduces transplacental transport of transthyretin and T4 to the placenta and developing fetus. This may contribute to the negative effects of smoking on fetal growth, development and pregnancy viability.


Genetic control of contact sensitivity to oxazolone in inbred, H-2 congenic and intra-H-2 recombinant strains of mice.
Delayed-type hypersensitivity (DTH) to 2-phenyl-4-ethoxymethylene-5-oxazolone (oxazolone) was found to be under multigenic control in inbred, H-2 congenic and intra-H-2 recombinant strains of mice. A high response was associated with haplotypes H-2d,a,k and low response with haplotype H-2b. DTH to oxazolone was high or intermediate in different F1 hybrids of high and low responder mice. In F2 and backcross generations a higher response was associated with the "dd", than with "bb" phenotype, while intermediate response was found in the heterozygote "db" mice. A study of H-2 recombinant strains suggests that a gene controlling the DTH response maps in the I-B subregion of the H-2 complex. The response was significantly modified by gene(s) which are not linked to the H-2 complex and have not been mapped. Since congenitally athymic nude (nu/nu) mice did not respond to oxazolone, this contact sensitivity is belived to be a T cell-dependent immune response.


Early At-Home Measurement of Adalimumab Concentrations to Guide Anti-TNF Precision Dosing: A Pilot Study.
Underdosing of adalimumab can result in non-response and poor disease control in patients with rheumatic disease or inflammatory bowel disease. In this pilot study we aimed to predict adalimumab concentrations with population pharmacokinetic model-based Bayesian forecasting early in therapy. Adalimumab pharmacokinetic models were identified with a literature search. A fit-for-purpose evaluation of the model was performed for rheumatologic and inflammatory bowel disease (IBD) patients with adalimumab peak (first dose) and trough samples (first and seventh dose) obtained by a volumetric absorptive microsampling technique. Steady state adalimumab concentrations were predicted after the first adalimumab administration. Predictive performance was calculated with mean prediction error (MPE) and normalised root mean square error (RMSE). Thirty-six patients (22 rheumatologic and 14 IBD) were analysed in our study. After stratification for absence of anti-adalimumab antibodies, the calculated MPE was -2.6% and normalised RMSE 24.0%. Concordance between predicted and measured adalimumab serum concentrations falling within or outside the therapeutic window was 75%. Three patients (8.3%) developed detectable concentrations of anti-adalimumab antibodies. This prospective study demonstrates that adalimumab concentrations at steady state can be predicted from early samples during the induction phase. The trial was registered in the Netherlands Trial Register with trial registry number NTR 7692 ( www.trialregister.nl ).


Metabolic engineering and mechanical investigation of enhanced plant autoluminescence.
The fungal bioluminescence pathway (FBP) was identified from glowing fungi, which releases self-sustained visible green luminescence. However, weak bioluminescence limits the potential application of the bioluminescence system. Here, we screened and characterized a C3'H1 (4-coumaroyl shikimate/quinate 3'-hydroxylase) gene from Brassica napus, which efficiently converts p-coumaroyl shikimate to caffeic acid and hispidin. Simultaneous expression of BnC3'H1 and NPGA (null-pigment mutant in A. nidulans) produces more caffeic acid and hispidin as the natural precursor of luciferin and significantly intensifies the original fungal bioluminescence pathway (oFBP). Thus, we successfully created enhanced FBP (eFBP) plants emitting 3 × 1011 photons/min/cm2 , sufficient to illuminate its surroundings and visualize words clearly in the dark. The glowing plants provide sustainable and bio-renewable illumination for the naked eyes, and manifest distinct responses to diverse environmental conditions via caffeic acid biosynthesis pathway. Importantly, we revealed that the biosynthesis of caffeic acid and hispidin in eFBP plants derived from the sugar pathway, and the inhibitors of the energy production system significantly reduced the luminescence signal rapidly from eFBP plants, suggesting that the FBP system coupled with the luciferin metabolic flux functions in an energy-driven way. These findings lay the groundwork for genetically creating stronger eFBP plants and developing more powerful biological tools with the FBP system.


Understanding the revenue cycle: coordinated billing process key to radiology success.
*An integrated approach across all involved departments is the key to revenue cycle success. *Accurate, complete, and timely data drives the revenue cycle. *Up-to-date and accurate coding and charge capture will help ensure that the institution is reimbursed correctly.


Stability of citrinin and deoxynivalenol during germination process of barley.
The stability of citrinin and deoxynivalenol during germination process of barley spiked with these mycotoxins at a level of 2/ig/g was investigated. Germinated barley was analyzed after 1, 3, and 5 days to follow the stability of citrinin and deoxynivalenol during the germination process. Two thin layer chromatographic methods were used for determinations of citrinin and deoxynivalenol. An average of 93.6% of citrinin and 77.1% of deoxynivalenol were destroyed within 5 days during the germination process of barley.


The model of the rheological behavior of gelatinized starch at low concentrations.
The optical density at the wavelength from 300 to 800 nm and the rheological behavior of gelatinized starch at its concentration from 3 to 30 g/L and the medium pH from 2 to 11 were investigated. The volume concentration of the swelling associates of macromolecules or their globules increases when pH of gelatinized starch rises. An influence of the volume concentration of gelatinized starch on the dependence of the shear stress on shear rate was studied. The dynamic viscosity is proportional to the volume concentration at the gelatinized starch concentration lower than 25% and proportional to the logarithm of the volume concentration at the starch concentration greater than 25%. Proposed mathematical model takes into account an influence of the volume concentration of swelling starch globules and the concentration of glucoside links of macromolecules between the globules of macromolecules or their associates on the dynamic viscosity of gelatinized starch. This model allows to describe the experimental data obtained.


Validation of phospholipase A2 receptor direct immunofluorescence staining in the diagnosis of primary membranous glomerulonephritis.
Distinguishing between primary and secondary subtypes of membranous glomerulonephritis (MGN) is critical for its clinical management. We prospectively compared direct immunofluorescence (DIF) staining for phospholipase A2 receptor (PLA2R) on frozen renal biopsy with the presence of detectable serum PLA2R antibody assessed by enzyme linked immunosorbent assay (ELISA) in the diagnosis of primary MGN. Forty-six patients with biopsy-proven MGN were enrolled from April 2017 to June 2019 with 31/46 (67.4%) being primary and 15/46 (32.6%) being secondary as determined by comprehensive clinical assessment. This is currently deemed to be the gold standard for distinguishing primary from secondary MGN. Amongst the 31 primary MGN patients, 24/31 were positive on PLA2R DIF staining compared to 18/31 being positive on the PLA2R ELISA (p=0.03). Amongst the 15 secondary MGN patients, 1/15 was positive on PLA2R DIF compared to 0/15 on PLA2R ELISA (p=1.0). In conclusion, the presence of PLA2R staining on DIF demonstrated superior sensitivity and similar specificity compared to the detection of circulating PLA2R antibodies by ELISA in the diagnosis of primary MGN in a cohort of 46 patients with biopsy-proven MGN. We suggest that DIF should be considered as part of routine work-up in all newly diagnosed cases of MGN.


Gait Retraining: Altering the Fingerprint of Gait.
In terms of running, there is evidence that links mechanics with injury. This evidence provides the justification for altering these mechanics. Increased hip adduction and vertical impact loading have been most commonly associated with injury. More work is needed in order to understand the optimal way to retrain gait patterns in runners. The human body has a considerable ability to adapt. To provide individuals with the ability to alter faulty movement patterns in ways that can reduce injury risk is a powerful tool.


Evaluating the 8-item overactive bladder questionnaire (OAB-v8) using item response theory.
The OAB-v8 is a patient-reported outcome questionnaire used to screen for overactive bladder and measure symptom bother. This study uses modern validation methods to assess the item and test characteristics of the OAB-v8, and determine whether it should be scored differently for men and women. A secondary analysis of data from patients with lower urinary tract symptoms prospectively recruited from a urology clinic in Calgary, Canada. Item-response theory (IRT) was used to evaluate the dimensionality, reliability, and validity of the OAB-v8, and differential functioning analysis (DIF) was used to determine whether the OAB-v8 should be scored differently for men and women. A total of 1128 patients were included for analysis, of which 28% were female. Mean OAB-v8 scores indicated meaningful levels of symptom bother, and women scored 2 points higher, on average, than men (P = 0.005). Reliability was high (Cronbach's alpha = 0.901) for a wide range of patients (between -2 and 2.5 standard deviations of the mean). Samejima's graded response model best fit the data, and the scale demonstrated essential unidimensionality. Items #5 and #6 were excessively correlated. There was no evidence of differential functioning. OAB-v8 demonstrated high reliability and would be appropriate for a wide range of patients. Items #5 and #6 may need to be re-worded. The OAB-v8 should not be scored differently for men and women, although women do appear to experience higher average levels of symptom bother from OAB than men.


Thermodynamic feasibility of shipboard conversion of marine plastics to blue diesel for self-powered ocean cleanup.
Collecting and removing ocean plastics can mitigate their environmental impacts; however, ocean cleanup will be a complex and energy-intensive operation that has not been fully evaluated. This work examines the thermodynamic feasibility and subsequent implications of hydrothermally converting this waste into a fuel to enable self-powered cleanup. A comprehensive probabilistic exergy analysis demonstrates that hydrothermal liquefaction has potential to generate sufficient energy to power both the process and the ship performing the cleanup. Self-powered cleanup reduces the number of roundtrips to port of a waste-laden ship, eliminating the need for fossil fuel use for most plastic concentrations. Several cleanup scenarios are modeled for the Great Pacific Garbage Patch (GPGP), corresponding to 230 t to 11,500 t of plastic removed yearly; the range corresponds to uncertainty in the surface concentration of plastics in the GPGP. Estimated cleanup times depends mainly on the number of booms that can be deployed in the GPGP without sacrificing collection efficiency. Self-powered cleanup may be a viable approach for removal of plastics from the ocean, and gaps in our understanding of GPGP characteristics should be addressed to reduce uncertainty.


Reconstruction of the shortened columella in mild bifid nose using a propeller flap based on the nasal columella artery.
Bifid nose generally appears short columellar and lacks the nasal tip. Here, we describe a surgical correction technique for correcting the short columellar and nasal tip of bifid nose using a local flap to discuss outcomes, patient selection, and complications based on 11 years of experience. Thirty-two patients with mild wide bifid nose and shortened columella were included in this retrospective study. All patients underwent nasal rhinoplasty using a propeller flap based on the nasal columella artery. Nasal columella length, horizontal distance of tip-defining point, and angle of facial convexity were evaluated based on three-dimensional simulation technology. Complications including possible underlying reasons were analyzed. Patient satisfaction was evaluated using a self-assessment survey. Nasal esthetics and function were considerably improved. Follow-up examinations during a period of 9 months on average demonstrated stable results. The columella length was 9.7 ± 4.6 mm preoperatively and 19.9 ± 3.2 mm postoperatively (P < 0.05). The horizontal distance of tip-defining point (mm) decreased to 18.9 ± 5.5 mm postoperatively from a preoperative 23.3 ± 5.4 mm (P < 0.05). There were no severe postoperative complications. Complications were scar, temporary hematoma, and mild infection of nasal skin. The majority of patients (97%) rated their outcome as improved and much improved. Surgical correction using a nasal columella artery propeller flap is an effective therapeutic approach for patients with mild bifid nose. The use of a local flap along with minimal donor-site morbidity and reliable outcomes contributes to the high patient acceptance. Meticulous surgical technique and careful patient selection are prerequisites for successful results.


Histologic and biomechanical evaluation of biologic meshes following colonization with Pseudomonas aeruginosa.
Biologic meshes have become increasingly popular for the repair of abdominal wall defects, especially in contaminated sites. The purpose of this study was to evaluate the histologic and biomechanical properties of biologic mesh in response to a bacterial encounter. A rat model of Pseudomonas aeruginosa colonization and infection of subcutaneously implanted biologic mesh was used. Samples of biologic meshes [acellular human dermis (ADM) and porcine small intestine submucosa (SIS)] were inoculated with P. aeruginosa (10(5) or 10(9) cfu) or saline as a control prior to wound closure (n = 6 per group). After 10 or 20 d, the meshes were harvested. The recovered meshes were analyzed for histologic changes and bacterial recovery as well as the material strength properties. Statistical significance (P < 0.05) was determined using 1-way analysis of variance or Mann-Whitney test. ADM and SIS colonized with 10(9) cfu P. aeruginosa showed an increased inflammatory response with an associated decrease in neo-vascularization (P < 0.05) at 20 d post-implantation compared with controls. P. aeruginosa had no effect on the tensile strength of ADM, but the tensile strength and modulus of elasticity were reduced for SIS compared with controls at 20 d. Bacterial colonization of ADM and SIS with 10(9)cfu P. aeruginosa negatively effected neovascularization and cellular re-population of the material over time but only SIS showed alterations in their biomechanical properties in response to this gram-negative bacterial challenge.


Hormones in wellness and disease prevention: common practices, current state of the evidence, and questions for the future.
Confusion and controversy surround the use of estrogen, progesterone, testosterone, growth hormone, and thyroid hormones. This article discusses age-related hormone loss and supplementation therapies for age-related hormonal deficiencies as possible first-line therapeutic modalities to be considered in our search to improve quality of life, prevent chronic illnesses, and maintain wellness. The well-informed use of hormones in wellness and disease prevention will result in symptomatic improvement and should be considered an integral part in the armamentarium of options we offer our patients.


Strategy for the detection and management of coronary artery disease. "Physiology before anatomy".
A strategy for the diagnosis of ischemic heart disease should be based on knowledge of the prevalence of the disease in population subgroups. Asymptomatic patients should not be routinely screened. Asymptomatic patients or patients with nonanginal chest pain should have both a positive exercise electrocardiogram and stress nuclear scan before a diagnosis of ischemic heart disease is justified or arteriography is recommended. Patients with atypical angina should be evaluated with exercise radionuclide ventriculography. Coronary arteriography is rarely needed for diagnosis and is most properly used as a preoperative evaluation of a patient who has symptoms uncontrolled by medical management, or in whom a significant amount of myocardium is at risk as determined by physiologic testing with exercise electrocardiography or stress nuclear techniques.


[Treatment strategies for traumatic anterior shoulder dislocation].
Anterior glenohumeral instability is the most frequent type of shoulder instability. This is often associated with labral and osseous lesions leading to recurrent instability. A detailed medical history, a physical examination and targeted diagnostic imaging are necessary to assess possible pathological soft tissue alterations as well as bony lesions of the humeral head and the glenoid bone. Early surgical treatment has been shown to reduce the risk of recurrence, especially in young active athletes, and can avoid secondary damage. Shoulder dislocations in older patients also require a detailed assessment and selection of treatment as persisting pain and limitation of movement can occur due to rotator cuff lesions and nerve injuries. The purpose of this article is to provide an overview of the currently available evidence and results regarding diagnostic considerations and conservative vs. surgical treatment and time to return to sport after treatment of a primary anterior shoulder dislocation.


Effect of nitric oxide donors and noradrenaline on Ca2+ release sites and global intracellular Ca2+ in myocytes from guinea-pig small mesenteric arteries.
In smooth muscle the spontaneous Ca2+ release from the sarcoplasmic reticulum (SR) occurs at preferred locations called frequent discharge sites (FDSs) giving rise to localized intracellular Ca2+ transients (Ca2+ sparks). Laser scanning confocal microscopy of fluo-3-loaded single myocytes freshly isolated from small mesenteric arteries of guinea-pig was used to investigate the action of nitric oxide (NO) donors and noradrenaline on the position and activity of FDSs and on global intracellular Ca2+ concentration ([Ca2+]i). In 8 % of cells 'microsparks', Ca2+ release events smaller in duration, spread and amplitude than Ca2+ sparks were observed. The location of the initiation point of Ca2+ sparks observed during line-scan imaging was found to 'jitter' by +/- 0.41 microm. However, the general position of an FDS within the cell did not change; most FDSs were close (within 1.2 +/- 0.1 microm) to the cell membrane and often multiple FDSs occurred in one confocal plane of the cell. In the resting state, NO donors S-nitroso-N-acetylpenicillamine (SNAP; 50 microM) and sodium nitroprusside (SNP; 100 microM) did not change the general position of FDSs and slightly depressed their activity, but did not affect the global [Ca2+]i significantly. Application of noradrenaline (1-10 microM) increased Ca2+ spark frequency at existing FDS(s) leading to a Ca2+ wave. The increase in FDS activity and in global [Ca2+]i produced by noradrenaline were inhibited by the presence of SNAP or SNP but not by 8-bromoguanosine cyclic 3',5'-monophosphate (8-Br-cGMP; 100 microM). In the presence of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), inhibitor of soluble guanylate cyclase, SNAP and SNP still exerted their effects on the noradrenaline response. These results suggest that SNAP and SNP inhibit the noradrenaline-evoked rise in global [Ca2+]i by a cGMP-independent mechanism and that part of this effect is due to inhibition of the activity of FDSs; moreover, only the activity, but not the position, of FDSs is changed by either stimulant or inhibitory substances.


Lesions of the caruncle: a clinicohistopathologic study.
This study reports 112 caruncular lesions seen in the Eye Pathology Laboratory of the Wilmer Institute during a 52-year period (1923--1976). These uncommon lesions are analyzed with regard to their relative occurrence and their clinical and histologic features. Clinical grouping of these lesions is presented to help in the differential diagnosis of caruncular lesions. Nevi, papillomas, and sebaceous gland hyperplasia were the most commonly encountered lesions, accounting for 65% of the total.


The novel neuropsychotropic agent milacemide is a specific enzyme-activated inhibitor of brain monoamine oxidase B.
The novel neuropsychotropic agent milacemide hydrochloride (2-n-pentylaminoacetamide HCl) is a highly selective substrate of the B form of monoamine oxidase (EC 188.8.131.52; MAO). Under the in vitro conditions used in the present study, milacemide acts as an enzyme-activated, partially reversible inhibitor of MAO-B. A reversible inhibition of MAO-A activity is also observed at high concentrations. The inhibitory activity of milacemide is significantly greater for MAO-B. In vivo, after single or repeated oral administration, a specific inhibition of MAO-B is apparent in brain and liver, with a lack of inhibition of the MAO-A activity. In contrast to the irreversible inhibitory action of L-deprenyl, the recovery of MAO-B activity in vivo after milacemide administration is significantly faster, a result suggesting that it is a partially reversible inhibitor. The selective inhibitory effect of milacemide for MAO-B in vivo is confirmed by its potentiation of phenylethylamine-induced stereotyped behavior, whereas vasopressor responses to tyramine were not affected. These observations suggest that milacemide could enhance dopaminergic activity in the brain and could be used as therapy for Parkinson's disease in association with L-3,4-dihydroxyphenylalanine.


The susceptibility of erythrocytes to oxidation during storage of blood: effects of melatonin and propofol.
We investigated the effects of melatonin and propofol in lipid peroxidation and antioxidant capacity of erythrocytes in stored bloods. Donated blood was taken into three citrate-phosphate-dextrose containing blood bags. One bag was used as control, the others were added either melatonin or propofol. Erythrocyte lipid peroxidation and antioxidant capacity and their sensitivity to in vitro oxidation were measured on days 0, 7, 14, 21 and 28. In control group, erythrocyte malondialdehyde levels and sensitivity to in vitro oxidation were increased whereas glutathione, glutathione peroxidase and superoxide dismutase levels were decreased. Melatonin prevented malondialdehyde accumulation and preserved glutathione, glutathione peroxidase and superoxide dismutase levels. Propofol preserved glutathione and glutathione peroxidase levels but did not affect catalase and superoxide dismutase activities. We showed that melatonin in stored blood could prevent lipid peroxidation and increase the resistance of erythrocytes to in vitro oxidation while propofol did not show such effects.


A comprehensive study concerning the reproducibility and stability of organic n-type field effect transistors is presented. C(60) based OFETs were chosen to investigate the fabrication reproducibility and the long term stability because C(60) is a high mobility n-type material. We fabricated 48 transistors and each transistor was measured for 24 h inside the glove box. To test for life time stability - long term measurements up to three months have been undertaken. We report about the fluctuations in the device parameters of all investigated transistors by comparing the transfer characteristics, and on/off ratio for short time and long time measurements. C(60) based OFETs showed good reproducibility and stability for short time measurements and a decay for long time measurements.


Birt-Hogg-Dubé syndrome detected incidentally by asymptomatic bilateral pneumothorax in health screening: a case of a young Japanese woman.
Birt-Hogg-Dubé syndrome (BHD) is an autosomal dominant disease caused by mutations of germline folliculin (FCLN) mapped in the chromosome 17p11.2 region. BHD commonly accompanies renal tumors, fibrofolliculomas, multiple pulmonary cysts, and spontaneous pneumothorax. We report a case of a young Japanese woman in whom asymptomatic bilateral pneumothorax was found incidentally in a health screening, which led to the diagnosis of BHD. She had developed neither renal tumors nor fibrofolliculomas. However, her father, uncle, and aunt also experienced pneumothorax. In Japan, BHD is not yet well known because skin-related symptoms of fibrofolliculomas are sometimes absent unlike in most cases in Europe and the United States. On the basis of this case, we propose that BHD should be considered at the time of pneumothorax examination.


Small airway disease in chronic obstructive pulmonary disease: insights and implications for the clinician.
Small airway disease (SAD) is a common feature in chronic obstructive pulmonary disease (COPD) patients. Chronic exposure to cigarette smoking causes inflammation, damage, tissue remodelling and eventually airway loss. These features lead to airflow limitation and defective alveolar ventilation. The aim of this review is to provide clinicians with an up-to-date perspective of SAD in COPD. In this review, we will discuss the key pathological features of SAD. We also review state-of-the-art techniques for measuring SAD, including impulse oscillometry and lung imaging methods. We discuss emerging concepts such as SAD in the earlier stages of COPD and the relationship between SAD and emphysema. The current review highlights the importance of targeting small airways early in the course of COPD to tackle disease progression.


Developing a special microwave oven: Assessment of its performance for dough fermentation and nutrient soup elaboration.
Bread and soup are two of the most important foods in daily life, thus dough fermentation and nutrient soup elaboration are more and more popular, but there is a lack of relevant low-cost and high-reliable household appliances on the market. Therefore, this paper proposes automatic control methods for dough fermentation and nutrient soup elaboration based on a special microwave oven. Fermentation theory, run-up microwave fermentation principle, microwave extraction principle, NTC temperature probe design and scalable fuzzy control algorithm are described in detail. Besides, the experimental platform is set up with a temperature chamber, an optical fiber thermometer and a power meter. Experimental results demonstrate that the relationship between the heating time and flour's mass is linear. For different ambient temperature tests, the volume ratios of the fermented dough to unfermented dough of different cases range from 2.2 to 2.62, and the inside of the dough after fermentation is fluffy, with small and dense cavities. Meanwhile, there is no acid taste and skin dryness, and the power consumption of microwave fermentation is less than half of that induced by grill, convection or steam fermentation. The detection error of the NTC temperature probe with microwave shielded is 0.48 °C, and the control error of the closed loop system is less than 0.5 °C. The temperature-rise slope of water is lower than that of ingredient, and the water's temperature is about 1 °C less than that of the ingredient. The soup after microwave elaboration is amber and clear, the ingredients are intact, the water loss is less than 50 g, and the total power consumption is 684 Wh. In short, microwave-based control methods for dough fermentation and nutrient soup elaboration are effective.


Mortality and cancer incidence in patients with dermatitis herpetiformis: a cohort study.
One hundred and fifty-two patients in whom a diagnosis of dermatitis herpetiformis was made at St John's Hospital for Diseases of the Skin, London, during 1950-85, were followed from the date of diagnosis to the end of 1989 for mortality, and from 1971, or the date of diagnosis if later, to 1986 for cancer incidence. Thirty-eight deaths occurred under the age of 85, slightly fewer than expected on the basis of national general population rates [standardized mortality ratio (SMR) = 87; 95% confidence interval (CI) 61-119]. All-cause mortality was somewhat lower in patients who had followed a gluten-free diet (SMR = 51; 17-120) than in those who had not (SMR = 97; 66-136), but the difference in SMRs was not significant (P = 0.3). Cancer mortality was non-significantly below expectations from national rates (SMR = 72; 31-142), but cancer incidence was significantly increased [standardized registration ratio (SRR) = 394; 180-749]. No particular cancer site accounted for the cancer incidence excess. One death occurred from cancer of the small intestine (SMR = 4953, P = 0.04), and one lymphoma was incident (SRR = 1555, P = 0.12). Increased risks of these malignancies have previously been found to be associated with coeliac disease (which is present in many patients with dermatitis herpetiformis), and with dermatitis herpetiformis, respectively. Mortality from ischaemic heart disease (IHD) was significantly below national rates (SMR = 37; 95% CI 12-86), and was similar in patients who had followed a gluten-free diet and those who had not.(ABSTRACT TRUNCATED AT 250 WORDS)


Synthesis and structure-activity studies of the V-ATPase inhibitor saliphenylhalamide (SaliPhe) and simplified analogs.
An efficient total synthesis of the potent V-ATPase inhibitor saliphenylhalamide (SaliPhe), a synthetic variant of the natural product salicylihalamide A (SaliA), has been accomplished aimed at facilitating the development of SaliPhe as an anticancer and antiviral agent. This new approach enabled facile access to derivatives for structure-activity relationship studies, leading to simplified analogs that maintain SaliPhe's biological properties. These studies will provide a solid foundation for the continued evaluation of SaliPhe and analogs as potential anticancer and antiviral agents.


Cerebral and cerebellar white matter tract alterations in patients with Pantothenate Kinase-Associated Neurodegeneration (PKAN).
To examine structural connectivity of white matter tracts in patients with Pantothenate Kinase-Associated Neurodegeneration (PKAN) dystonia and identify those ones which correlate negatively to severity of symptoms. In a group of 41 patients suffering from PKAN dystonia and an age- and gender-matched control group, white matter tractography was carried out, based on diffusion tensor imaging magnetic resonance data. Postprocessing included assessment of Quantitative Anisotropy (QA) using q-space diffeomorphic reconstruction in order to reduce influence of iron accumulation in globus pallidus of patients. Whole brain tractography presented significantly reduced QA values in patients (0.282 ± 0.056, as compared to controls (0.325 ± 0.046, p < 0.001). 9 fiber clusters of tracts correlated negatively to the dystonia score of patients: the middle cerebellar peduncle and the tracts of both cerebellar hemispheres as well as corpus callosum, forceps minor, the superior cortico-striate tracts and the superior thalamic radiations of both cerebral hemispheres (False Discovery Rate FDR = 0.041). The finding of a reduced global structural connectivity within the white matter and of negative correlation of motor system-related tracts, mainly those between the basal ganglia, cortical areas and the cerebellum, fits well to the concept of a general functional disturbance of the motor system in PKAN.


ppBAM: ProteinPaint BAM track for read alignment visualization and variant genotyping.
ProteinPaint BAM track (ppBAM) is designed to assist variant review for cancer research and clinical genomics. With performant server-side computing and rendering, ppBAM supports on-the-fly variant genotyping of thousands of reads using Smith-Waterman alignment. To better visualize support for complex variants, reads are realigned against the mutated reference sequence using ClustalO. ppBAM also supports the BAM slicing API of the NCI Genomic Data Commons (GDC) portal, letting researchers conveniently examine genomic details of vast amounts of cancer sequencing data and reinterpret variant calls. BAM track examples, tutorial, and GDC file access links are available at https://proteinpaint.stjude.org/bam/. Source code is available at https://github.com/stjude/proteinpaint.


Comparison of In-Hospital Outcomes and Readmission Rates of Transcatheter Aortic Valve Implantation in Mixed Aortic Valve Disease Versus Pure Aortic Stenosis.
Recently, transcatheter aortic valve implantation (TAVI) has been performed in patients with combined aortic stenosis (AS) and aortic regurgitation. We sought to evaluate in-hospital outcomes and readmission rates after TAVI in patients with mixed aortic valve disease (MAVD). A total of 100,573 TAVI procedures were identified between 2011 and 2017 using International Classification of Diseases, Ninth Revision and International Classification of Diseases, Tenth Revision procedure codes the from Nationwide Readmissions Database. We separated patients into 2 cohorts, those with MAVD and those with pure AS. The primary outcome was all-cause inpatient mortality after TAVI, and secondary outcomes included rates of 30- and 90-day readmissions and postprocedural complications. A total of 3,260 patients had MAVD (median age 83 years, 43.5% women). In-hospital mortality (2.5% vs 2.6%, p = 0.531) and rates of paravalvular leak (1.0% vs 1.3%, p = 0.056) were similar between the MAVD and pure AS groups. Major bleeding (7.4% vs 9.6%, p <0.001), 30-day readmission (0.5% vs 8.8%, p <0.001) and 90-day readmission rates (0.8% vs 16.0%, p <0.001), acute kidney injury (12.9% vs 15.1%, p <0.001), postoperative ischemic stroke (2.0% vs 5.7%, p <0.001), and mechanic circulatory support use (1.9% vs 4.5%, p <0.001) were less prevalent in the MAVD cohort. Using a multivariate logistic regression model to adjust for confounding factors, MAVD was not predictive of mortality in patients who underwent TAVI (adjusted odds ratio [adjOR] 1.25, 95% confidence interval [CI] 0.99 to 1.57, p = 0.056); however, MAVD was associated with: decreased odds of 30-day readmission (adjOR 0.05, 95% CI 0.03 to 0.08, p <0.001), 90-day readmission rates (adjOR 0.04, 95% CI 0.03 to 0.06, p <0.001), and higher odds of pacemaker implantation (adjOR 1.46, 95% CI 1.29 to 1.65, p <0.001). In conclusion, despite differences in the aortic valve and left ventricular anatomy (pressure vs volume-related adaptive changes) in patients with MAVD and pure AS, TAVI appears safe and feasible. However, patients with MAVD were more likely to have permanent pacemakers implanted. The results of our study warrant further randomized controlled studies to confirm these findings.


Harvest technique for pedicled transposition of latissimus dorsi muscle: an old trade revisited.
Transposition of extrathoracic muscle flaps has been the cornerstone of treatment of a number of complex intrathoracic pathologies such as bronchopleural fistulas and residual infected pleural spaces. We present a simple step-wise technique for preservation and harvesting of the most common muscle flap employed by thoracic surgeons, namely latissimus dorsi, just prior to performing a standard posterolateral thoracotomy. Since 2004, we have successfully utilized pedicled latissimus muscle as our preferred prophylactic flap against development of postoperative bronchopleural fistulas or recurrent empyemas. This technique should be part of every thoracic surgeon's surgical armamentarium.


Tofacitinib Reduces Spinal Inflammation in Vertebral Bodies and Posterolateral Elements in Ankylosing Spondylitis: Results from a Phase 2 Trial.
This post hoc analysis of phase 2 trial data assessed the efficacy of tofacitinib on magnetic resonance imaging (MRI) outcomes with the detailed anatomy-based Canada-Denmark (CANDEN) MRI scoring system and evaluated tofacitinib suppression of spinal inflammation in patients with active ankylosing spondylitis (AS). Patients with active AS (per modified New York criteria) were randomized 1:1:1:1 to receive tofacitinib 2, 5, or 10 mg twice daily (BID), or placebo, in a 16-week, phase 2, double-blind clinical trial. Spine MRI assessments were performed at baseline and week 12. For post hoc analysis, MRI images from patients receiving tofacitinib 5 or 10 mg BID, or placebo, were re-evaluated by two readers blinded to time point/treatment and assessed by the CANDEN MRI scoring system. Least squares mean changes from baseline to week 12 were reported for CANDEN-specific MRI outcomes, with analysis of covariance used for comparisons of pooled tofacitinib and tofacitinib 5 or 10 mg BID versus placebo. p values without multiplicity adjustment were reported. MRI data from 137 patients were analyzed. At week 12, CANDEN spine inflammation score and vertebral body, posterior elements, corner, non-corner, facet joint, and posterolateral inflammation subscores were significantly reduced with pooled tofacitinib versus placebo (p < 0.0001; except non-corner subscore, p < 0.05). Total spine fat score was numerically increased with pooled tofacitinib versus placebo. In patients with AS, tofacitinib treatment was associated with significant reductions in MRI scores of spinal inflammation versus placebo, as assessed by the CANDEN MRI scoring system. Tofacitinib reduced inflammation in posterolateral elements of the spine and facet joints, which has not been described previously. ClinicalTrials.gov registry (NCT01786668).


Teaching the pelvic organ prolapse quantitation system.
The objective of this study was to determine the ease with which the new Pelvic Organ Prolapse Quantitation system could be learned by residents and medical students. Standardized multiple-choice tests were administered to 51 obstetric and gynecology residents and medical students from 4 community-based and university-based programs. Parallel pretests and posttests were administered in conjunction with a 17-minute video demonstration of the Pelvic Organ Prolapse Quantitation system and with the addition of a visual memory aid. The posttest was repeated 3 months after the video presentation. The use of a 17-minute video significantly enhanced participants' ability to interpret examination findings when expressed in the terminology of the system (mean improvement in scores 33%, P <.0001). Posttest scores were similar regardless of the type of program or exposure to urogynecology faculty. These scores were maintained at the 3-month retesting. The Pelvic Organ Prolapse Quantitation system can be effectively taught by means of a public-domain video presentation.


Hydrophobic binding of the ectodomain of influenza hemagglutinin to membranes occurs through the "fusion peptide".
Toward elucidating molecular details of virus-induced membrane fusion, we have studied the low pH-triggered interaction of the bromelain-solubilized ectodomain of influenza hemagglutinin with liposomes. Polypeptide segments which insert into the apolar phase of the lipid bilayer were first labeled specifically using either of the two membrane-restricted carbene-generating reagents, 3-(trifluoromethyl)-3-([125I]iodophenyl)diazirine and 1-palmitoyl-2-[11-[4-[3-(trifluoromethyl)diazirinyl]phenyl] undecanoyl]-sn-glycero-3-phosphorylcholine, and were then identified on the basis of cyanogen bromide and 2-(2-nitrophenylsulfenyl)-3-methyl-3'-bromoindolenine-skatole fragment analysis and Edman degradations. Here, we demonstrate that the hydrophobic interaction is mediated solely by the so-called "fusion peptide" which corresponds to the NH2-terminal segment of the BHA2 subunit of nature influenza hemagglutinin. Predominant sites of labeling within that segment were Phe-3, Ile-6, Phe-9, Trp-14, Met-17, and Trp-21. The average 3-4 residue spacing between consecutive labeled amino acid side chains suggests a helical structure of that segment with an amphiphilic character.


Persistent Arthralgia Associated with Chikungunya Virus Outbreak, US Virgin Islands, December 2014-February 2016.
After the 2014-2015 outbreak of chikungunya virus in the US Virgin Islands, we compared the prevalence of persistent arthralgia among case-patients and controls. Prevalence was higher in case-patients than controls 6 and 12 months after disease onset. Continued vaccine research to prevent acute illness and long-term sequelae is essential.


DRiPs solidify: progress in understanding endogenous MHC class I antigen processing.
Defective ribosomal products (DRiPs) are a subset of rapidly degraded polypeptides that provide peptide ligands for major histocompatibility complex (MHC) class I molecules. Here, recent progress in understanding DRiP biogenesis is reviewed. These findings place DRiPs at the center of the MHC class I antigen processing pathway, linking immunosurveillance of viruses and tumors to mechanisms of specialized translation and cellular compartmentalization. DRiPs enable the immune system to rapidly detect alterations in cellular gene expression with great sensitivity.


Synergistic cancer immunotherapy combines MVA-CD40L induced innate and adaptive immunity with tumor targeting antibodies.
Virus-based vaccines and appropriate costimulation potently enhance antigen-specific T cell immunity against cancer. Here we report the use of recombinant modified vaccinia virus Ankara (rMVA) encoding costimulatory CD40L against solid tumors. Therapeutic treatment with rMVA-CD40L-expressing tumor-associated antigens results in the control of established tumors. The expansion of tumor-specific cytotoxic CD8+ T cells is essential for the therapeutic antitumor effects. Strikingly, rMVA-CD40L also induces strong natural killer (NK) cell activation and expansion. Moreover, the combination of rMVA-CD40L and tumor-targeting antibodies results in increased therapeutic antitumor efficacy relying on the presence of Fc receptor and NK cells. We describe a translationally relevant therapeutic synergy between systemic viral vaccination and CD40L costimulation. We show strengthened antitumor immune responses when both rMVA-CD40L-induced innate and adaptive immune mechanisms are exploited by combination with tumor-targeting antibodies. This immunotherapeutic approach could translate into clinical cancer therapies where tumor-targeting antibodies are employed.


Left Ventricular Systolic Function After 3 Months of SGLT2 Inhibitor Therapy in Heart Failure Patients with Reduced Ejection Fraction.
Not much is known about the effects of sodium-glucose cotransporter 2 inhibitors (SGLT2i) on echocardiographic parameters of left ventricular (LV) systolic function in patients with heart failure and reduced ejection fraction (HFrEF).We prospectively included 59 outpatients with HFrEF: 41 patients received SGLT2i with OMT (SGLT2i+ group), whereas eighteen patients received OMT without SGLT2i (SGLT2i- group). Myocardial work index (MWI), 3D ejection fraction (3D LVEF), and global longitudinal strain (GLS) were measured at baseline and after 3 months following treatment. At 3-month follow-up, the SGLT2i+ group showed significantly greater improvement in MWI than the SGLT2i- group. In both groups, there was a significant improvement in 3D LVEF and LV GLS, circulating NT-proBNP levels, and NYHA functional class, with significantly greater improvement in the SGLT2i+ group.In conclusion, the addition of SGLT2i to fully optimized background medical therapy resulted in a greater improvement of LV systolic function among outpatients with HFrEF.


Management of medication-induced gingival hyperplasia: a systematic review.
Medication-induced gingival hyperplasia (MIGH) has been linked to several medications, with a reported prevalence ranging between 0.5% and 85%. The aim of this study was to systematically review the management approaches for MIGH and estimate recurrence rate and time to relapse. An electronic literature search was conducted using PICO questions (P = patients with medication-induced gingival hyperplasia; I = surgical and/or nonsurgical treatment options; C = no control is required; and O = partial or complete resolution and recurrence) and medical subject heading terms in the PubMed and Web of Science databases following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses protocol up to December 2019. All English-language articles on MIGH surgical and nonsurgical management options were included. Eligible articles were systematically reviewed and assessed for bias using preset criteria and multiple levels of elimination. Data were extracted from eligible studies and analyzed. Twenty-two eligible articles were included in this study. Management approaches included discontinuation or change of the offending medication if medically feasible in addition to surgical and nonsurgical interventions. Nonsurgical approach included scaling and root planing, oral hygiene instructions, and antimicrobial mouthrinses. Persistent or relapsed cases had complete resolution with excision of hyperplastic gingiva. Laser-assisted surgeries combined with intensive plaque control measures demonstrated less risk of recurrence. Several treatment options for MIGH have been reported with variable outcomes. Duration and size of hyperplastic gingival tissue may have an effect on overall recurrence rate.


Rationale and design of the Adapted Physical Activity in advanced Pancreatic Cancer patients (APACaP) GERCOR (Groupe Coopérateur Multidisciplinaire en Oncologie) trial: study protocol for a randomized controlled trial.
Exercise during chemotherapy is a promising strategy to reduce fatigue and improve health-related quality of life (HRQoL). It has been shown to be feasible and efficient in various cancers, including advanced-stage cancers. Effects of physical activity have never been explored in advanced pancreatic ductal adenocarcinoma (PDAC). We aim to evaluate the effects of an exercise intervention in this setting. This randomized, national, multicenter, interventional study will examine the effectiveness of an unsupervised, home-based, 16-week adapted physical activity (APA) program. Specificities of advanced PDAC for the implementation of the APA program will be taken into account (healthy volunteer as physical activity partner instead of patient groups, nutritional management). The main inclusion criteria are: patients with histologically confirmed, unresectable PDAC; scheduled for chemotherapy; performance status 0-2; age ≥ 18; and physical activity partner. In total, 200 patients will be randomized into either the APA program (aerobic and resistance exercises) in addition to usual care (including chemotherapy at the investigator's choice), or usual care. The primary objective will be the effect on fatigue (Multidimensional Fatigue Inventory, MFI-20) and HRQoL (European Organization for Research and Treatment of Cancer-Quality of Life-C30 questionnaire, EORTC-QLQ-C30; co-primary endpoint) at week 16. As secondary objectives, the effects of the exercise intervention on pain, anxiety, depression, nutritional status, insulin resistance, tolerance of chemotherapy, survival, and adherence to the APA program will be evaluated. Patients with advanced PDAC are strongly affected by fatigue, and are thus likely to benefit from an exercise intervention. Moreover, exercise may have a potential beneficial effect on tumor outcome by reducing insulin resistance and insulin/insulin-like growth factor-1 (IGF-1) secretion. However, an exercise intervention may appear challenging due to multiple PDAC-related symptoms such as fatigue, depression, pain, and denutrition. We hypothesize that an APA program taking into account specific characteristics of PDAC may improve symptoms and HRQoL. If demonstrated to be feasible and effective, such APA programs will be systematically proposed to patients with advanced PDAC in addition to usual care. ClinicalTrial.gov NCT02184663 ; Registration date: 2 July 2014.


The pattern of change in the lecithin/sphingomyelin ratio in patients with preterm premature rupture of membranes between 24 and 34 weeks' gestation.
To report the pattern of change in the lecithin/sphingomyelin (L/S) ratio in patients with preterm premature rupture of membranes (PPROM) between 24 and 34 weeks' gestation. L/S was determined prospectively using transvaginally and transabdominally collected amniotic fluid from patients with PPROM between 24 and 34 weeks' gestation. Samples were collected prospectively on admission and every 48 to 96 hours until L/S was > or =2.0. All patients received intramuscular betamethasone weekly. Fifty-five patients were included in the study. One hundred twenty-seven samples were collected transvaginally and nine were collected transabdominally. Cox regression analysis showed that a higher initial L/S value and more advanced gestational age were associated with accelerated lung maturation. Among patients at > or =29 weeks' gestation with an initial L/S of > or =1.5 and <2.0 (n=17), 15 of 17 (88%) reached L/S > or = 2 at a mean of 3.1 +/- 1.7 days (range 1.0 to 7.0 days). With an initial L/S of > or =1.0 and <1.5 (n=16), 14 of 16 (88%) patients reached L/S > or =2 at a mean of 4.1 +/- 1.9 days (range 1.7 to 7.0 days). With an initial L/S of <1.0 (n=11), 6 of 11 (54%) patients reached L/S > or =2 at a mean of 5.0 +/- 1.6 days (range 4.7 to 6.8 days). Our data document a dramatic acceleration of fetal lung maturation among patients with PPROM at > or = 29 weeks.


The road to diagnosis and treatment in girls and boys with ADHD - gender differences in the diagnostic process.
The number of referrals for diagnostic assessments of Attention Deficit/Hyperactivity Disorder (ADHD) has increased in the last decade. There is a lack of studies examining the diagnostic process and the treatment provided, particularly from a gender perspective. From a consecutive cohort of Child and Adolescent Psychiatric (CAP) outpatients, the medical records of 50 boys and 50 girls (under 18 years of age) with a diagnosis of ADHD were selected by an Excel random numbers generator. Data about referral reason, diagnostic process and treatment were analysed. Emotional symptoms were more common as a reason for referral to CAP among girls, whereas neurodevelopmental disorders were more common among boys. Compared to the boys, the girls were older at first visit to CAP and at the ADHD diagnosis. The girls had had more visits to the clinic prior to the ADHD diagnostic decision and had more often been prescribed non-ADHD medication both before and after the ADHD diagnosis. The rate of ADHD medication was similar in boys and girls. Girls had more often been admitted to a CAP inpatient care unit prior to the ADHD diagnosis due to acute psychiatric symptoms, and had received more individual psychotherapeutic counselling. The results highlight the need for broader psychiatric investigations including neuropsychiatric symptoms in girls referred for 'emotional problems'.


Multi-color live-cell fluorescence imaging of primary ciliary membrane assembly and dynamics.
The ciliary membrane is continuous with the plasma membrane but has distinct lipid and protein composition, which is key to defining the function of the primary cilium. Ciliary membranes dynamically assemble and disassemble in association with the cell cycle and directly transmit signals and molecules through budding membranes. Various imaging approaches have greatly advanced the understanding of the ciliary membrane function. In particular, fluorescence live-cell imaging has revealed important insights into the dynamics of ciliary membrane assembly by monitoring the changes of fluorescent-tagged ciliary proteins. Protein dynamics can be tracked simultaneously using multi-color live cell imaging by coupling ciliary-associated factors with different colored fluorescent tags. Ciliary membrane and membrane associated-proteins such as Smoothened, 5-HTr6, SSTR3, Rab8a, and Arl13b have been used to track ciliary membranes and centriole proteins like Centrin1/2, CEP164, and CEP83 are often used to mark the ciliary basal body. Here, we describe a method for studying ciliogenesis membrane dynamics using spinning disk confocal live-cell imaging.


Interaction of a viral insulin-like peptide with the IGF-1 receptor produces a natural antagonist.
Lymphocystis disease virus-1 (LCDV-1) and several other Iridoviridae encode viral insulin/IGF-1 like peptides (VILPs) with high homology to human insulin and IGFs. Here we show that while single-chain (sc) and double-chain (dc) LCDV1-VILPs have very low affinity for the insulin receptor, scLCDV1-VILP has high affinity for IGF1R where it can antagonize human IGF-1 signaling, without altering insulin signaling. Consequently, scLCDV1-VILP inhibits IGF-1 induced cell proliferation and growth hormone/IGF-1 induced growth of mice in vivo. Cryo-electron microscopy reveals that scLCDV1-VILP engages IGF1R in a unique manner, inducing changes in IGF1R conformation that led to separation, rather than juxtaposition, of the transmembrane segments and hence inactivation of the receptor. Thus, scLCDV1-VILP is a natural peptide with specific antagonist properties on IGF1R signaling and may provide a new tool to guide development of hormonal analogues to treat cancers or metabolic disorders sensitive to IGF-1 without affecting glucose metabolism.


Chondrocyte-seeded collagen matrices implanted in a chondral defect in a canine model.
The objective of our study was to evaluate reparative tissues formed in chondral defects in an adult canine model implanted with cultured autologous articular chondrocytes seeded in type I and II collagen GAG matrices. Two defects were produced in the trochlea grooves of the knees of 21 dogs, with cartilage removed down to the tidemark. This study includes the evaluation of 36 defects distributed among five treatment groups: Group A, type II collagen matrix seeded with autologous chondrocytes under a sutured type II collagen flap; Group B, type I collagen matrices seeded with chondrocytes under a sutured fascia flap; Group C, unseeded type I collagen matrix implanted under a sutured fascia flap; Group D, fascia lata flap alone; and Group E, untreated defects. All animals were killed 15 weeks after implantation. Six other defects were created at the time of death and evaluated immediately after production as 'acute defect controls'. In three additional defects, unseeded matrices were sutured to the defect and the knee closed and reopened after 30 min to determine if early displacement of the graft was occurring; these defects served as 'acute implant controls'. The areal percentages of four tissue types in the chondral zone of the original defect were determined histomorphometrically: fibrous tissue (FT); hyaline cartilage (HC); transitional tissue (TT, including fibrocartilage); and articular cartilage (AC). New tissue formed in the remodeling subchondral bone underlying certain defects was also assessed. Bonding of the repair tissue to the subchondral plate and adjacent cartilage, and degradation of the adjacent tissues were evaluated. There were no significant differences in the tissues filling the original defect area of the sites treated with chondrocyte-seeded type I and type II matrices. Most of the tissue in the area of the original defect in all of the groups was FT and TT. The areal percentage of HC plus AC was highest in group E, with little such tissue in the cell-seeded groups, and none in groups C and D. The greatest total amount of reparative tissue, however, was found in the cell-seeded type II matrix group. Moreover, examination of the reparative tissue formed in the subchondral region of defects treated with the chondrocyte-seeded collagen matrices (Groups A and B) demonstrated that the majority of the tissue was positive for type II collagen and stained with safranin O. These results indicate an influence of the exogenous chondrocytes on the process of chondrogenesis in this site. In all groups with implants (A-D), 30(50% of the FT and TT was bonded to the adjacent cartilage. Little of this tissue (6-22%) was attached to the subchondral plate, which was only about 50% intact. Remarkable suture damage was found in sections from each group in which sutures were used. Harvest sites showed no regeneration of normal articular cartilage, 18 weeks after the biopsy procedure. Future studies need to investigate other matrix characteristics, and the effects of cell density and incubation of the seeded sponges prior to implantation on the regenerative response.


Cophylogeny and convergence shape holobiont evolution in sponge-microbe symbioses.
Symbiotic microbial communities of sponges serve critical functions that have shaped the evolution of reef ecosystems since their origins. Symbiont abundance varies tremendously among sponges, with many species classified as either low microbial abundance (LMA) or high microbial abundance (HMA), but the evolutionary dynamics of these symbiotic states remain unknown. This study examines the LMA/HMA dichotomy across an exhaustive sampling of Caribbean sponge biodiversity and predicts that the LMA symbiotic state is the ancestral state among sponges. Conversely, HMA symbioses, consisting of more specialized microorganisms, have evolved multiple times by recruiting similar assemblages, mostly since the rise of scleractinian-dominated reefs. Additionally, HMA symbioses show stronger signals of phylosymbiosis and cophylogeny, consistent with stronger co-evolutionary interaction in these complex holobionts. These results indicate that HMA holobionts are characterized by increased endemism, metabolic dependence and chemical defences. The selective forces driving these patterns may include the concurrent increase in dissolved organic matter in reef ecosystems or the diversification of spongivorous fishes.


Amniocentesis carried out for neural tube indications in South Wales 1973-1981: outcome of pregnancies and findings in the malformed abortuses.
The 2872 second trimester amniocentesis followed by amniotic alphafetoprotein (AFP) estimations carried out in South Wales between 1973 and 1981 on women known to be at increased risk for neural tube defect (NTD) and those who had a raised serum AFP level in an NTD screening programme led to the identification of 78 pregnancies of a fetus with anencephalus, 61 with 'open' spina bifida, 8 with gastroschisis, 3 with exomphalos, 2 with encephalocele and 6 with chromosome abnormality. Pregnancies of fetuses having 4 potentially identifiable NTDs were missed because of an equivocal AFP level and there were two false positive results leading to the termination of one normal fetus. It is emphasized that both the latter problems of one normal fetus. It is emphasized that both the latter problems would not have occurred had gel-electrophoresis for isoenzymes of acetyl cholinesterase been available. Follow-up of pregnancies showed that 7 children with 'closed' NTD and 3 with congenital hydrocephalus were born. The anencephalic and the 'open' spina bifida had a more florid lesion than is usual at term. Nearly all the spina bifida were associated with hydrocephalus, often severe and with an obvious Arnold-Chiari malformation. All but 13 had leg or back deformation or malformations in other systems, mostly in the renal tract.


Clinical considerations of bleomycin based electrochemotherapy with variable electrode geometry electrodes for inoperable, deep-seated soft tissue sarcomas.
The aim of the current prospective pilot study exclusively for deep-seated soft tissue sarcomas (STS) was to evaluate efficacy and safety of bleomycin-based ECT using VEG (variable electrode geometry) electrodes. During a 2-year period, seven surgically inoperable STSs were treated at the University of Szeged, Department of Surgery in Hungary. Electrode placement was determined by software planning using preoperative imaging (CT/MRI) and intraoperative ultrasound. Intravenous bleomycin (15.000 IU/m2) was administered 8 min before first pulse generation which lasted up to 40 min. Tumour response was evaluated through CT/MRI 2 months after treatment as per RECIST v.1.1. Five male- and 2 female patients were treated with fibromyxoid sarcoma (n = 2), epitheloid sarcoma (n = 3), liposarcoma (n = 1) and myofibroblastic sarcoma (n = 1) with median age of 54 years (49-88). Median tumour diameter, tumour volume and tumour depth was 5.9 cm (3.7-22.5), 131.13 cm3 (35.6-2456.22) and 6.18 cm (3.74-18.18), respectively. Median operative time was 75 min (35-180), median hospital stay 2 days (2-20). Two month follow-up confirmed partial response in 5 patients, while stable disease in 1 patient, and progressive disease in 1 case as per RECIST v.1.1. Grade 2 ulceration was experienced in four cases, and a transient left musculus quadriceps femoris plegia occured in one patient. Local control of deep-seated STSs with BLM-based VEG ECT holds a promising perspective and our results may serve as a practical guide for further investigation and treatment planning.


Post-Acute Care for Traumatic Brain Injury Patients in Taiwan.
Traumatic brain injury (TBI) can result in functional impairments. Many patients with TBI require post-acute care to improve their functional skills and allow for greater self-independence and a better quality of life. Taiwan's National Health Insurance proposed a nationwide post-acute care program in 2017 for patients with traumatic brain injury. The program's goal was to maximize patients' functional recovery and make it possible for them to return to their homes and communities. This study aimed to explore the effectiveness of such programs in Taiwan. This pilot study retrospectively evaluated a de-identified database that contained the scores of various assessments evaluated at admission and discharge. It used the data to determine the functional improvement of patients with traumatic brain injury after participating in post-acute care programs. This study collected complete data from 27 patients. After an average of 45.11 days in the program, the patients showed significant improvement in the Barthel Activity Daily Living Index, the Lawton-Brody Instrumental Activity Daily Living Scale, the Mini Nutrition Assessment, the EuroQol Five Dimensions Questionnaire, the Berg Balance Scale, the Galveston Orientation and Amnesia Test, the Rancho Los Amigos Scale, the Concise Chinese Aphasia Test, and the Mini Mental State Examination. After discharge, 96% of the patients could return to their community. This pilot study concluded that the program significantly promoted functional recovery for patients and could help patients with traumatic brain injury return to their communities and reduce the risk of institutionalization. Thus, patients with the potential for functional recovery could receive post-acute care in regional or community hospitals immediately after being discharged from acute wards. In the future randomized controlled trials are needed to further confirm the clinical impact of this program.


Tuberculosis outbreak in a long-term care facility.
Tuberculosis (TB) was a major cause of morbidity and mortality in Canada early in the 20th century. Elderly populations in Canada remain at high risk for TB disease. Elderly patients may present atypically, with the result that many active cases can remain undiagnosed. We present an outbreak of TB that occurred in a Residential and Long-Term Care (LTC) facility in Ontario. Case finding was carried out through the conventional concentric circle approach. Three rounds of tuberculin skin testing were conducted at 8-12 week intervals. Laboratory analysis was conducted at the Public Health Ontario Laboratories. An indoor air quality assessment was conducted to determine whether inadequate engineering controls were a transmission risk factor. A case of active pulmonary TB was confirmed in May 2010 in a staff member at the facility. By January 2011, 3 additional active cases and 24 latent tuberculosis infections among residents and staff had been identified. Genotyping methods confirmed that the 4 active cases were infected by an identical strain. Nine of 15 locations tested in the facility had air exchange rates below published guidelines. Prompt reporting of the initial case allowed for a quick initiation of the epidemiologic investigation. Given the epidemiology of TB in elderly populations, outbreaks should remain a concern for LTC facilities and physicians, even in jurisdictions of low TB incidence. Baseline and annual TB screening for residents and staff, early diagnosis of active TB, and adequate ventilation are important to reduce the incidence of disease.


Activation of K cells in mice with transplanted tumours differing in immunogenicity and metastasizing capacity.
The effector arm of antibody-dependent cellular cytotoxicity (ADCC) was evaluated using 51Cr-labelled chicken erythrocytes as targets in BALB/c mice transplanted with the Moloney sarcoma virus-induced tumours T-MSV and MS2, and in C57BL/6 mice transplanted with the chemically induced FS6 sarcoma, Lewis lung carcinoma and B16 melanoma. Tumour-bearing animals showed higher levels of ADCC than normal mice, a stimulation confirmed in MS2-bearing mice, using SL2 lymphoma cells as targets in a cytostasis assay. ADCC effector-cell capacity was higher in animals transplanted with the immunogenic, spontaneously regressing T-MSV than in mice bearing the poorly immunogenic metastasizing MS2 sarcoma. The increased ADCC activity detectable in the spleen of tumour-bearing hosts was not abolished by removal of phagocytic-adherent cells.


Dosimetry in isolation perfusion of the limbs by assessment of perfused tissue volume and grading of toxic tissue reactions.
The optimal single dosage of melphalan in isolation perfusion of the limbs for malignant melanoma was assessed. For this purpose a method to determine the volume of the isolated region in the individual patient and a grading system for the reaction of the normal tissues were introduced. A strictly standardized pharmacosurgical routine was developed that permitted an analysis of the correlation between dosage and the grade of toxic reaction in 90 perfusions. The optimal dosage of a cytostatic drug was considered to be the highest amount tolerated at an acceptable risk. Melphalan at 10 mg/l perfused tissue was determined as the likely optimum. This dose provoked remarkably little variation in toxicity, all reactions falling within a safe range. No exception to the applicability of this dosage was encountered.


NADPH oxidase 4 contributes to connective tissue growth factor expression through Smad3-dependent signaling pathway.
Transforming growth factor-β (TGF-β)/Smad signaling has been implicated in connective tissue growth factor (CTGF) expression in vascular smooth muscle cells (VSMC). Reactive oxygen species (ROS) are involved in activation of TGF-β/Smad signaling. However, detailed mechanisms underlying the process remain unclear. In present study, we demonstrated TGF-β1 strongly induced CTGF expression, Smad3 activation, NADPH oxidase 4 (Nox4) expression and increased ROS production in primary rat VSMC in vitro. NADPH oxidases inhibitor diphenylene iodonium (DPI) eliminated TGF-β1-induced CTGF expression and ROS generation. In addition, small-interfering RNA (siRNA) silencing of Smad3 or Nox4 significantly suppressed TGF-β1-mediated CTGF expression in VSMC. Furthermore, Nox4 silencing or inhibition eliminated TGF-β1-induced Smad3 activation and interaction between Nox4 and Smad3. In vivo studies further identified a positive correlation of Nox4 levels with Smad3 activation and CTGF expression in atherosclerotic arteries of patients and animal models. These data established that a novel mechanistic link of Nox4-dependent activation of Smad3 to increased TGF-β1-induced CTGF in the process of vascular remodeling, which suggested a new potential pathway for therapeutic interventions.


Comparison of IL-10 levels in chronic venous insufficiency ulcers and autologous donor tissue.
In previous immunohistochemical studies, chronic venous insufficiency (CVI) ulcers have been shown to display positive staining for interleukin-10 (IL-10), while other wounds (including autologous donor wound tissue) show a reduced staining level. IL-10 inhibits the synthesis of many proinflammatory cytokines, while also inhibiting antigen presentation by antigen-presenting cells. It is possible that abnormally high amounts of IL-10 in chronic wounds may be related to the failure of these wounds to progress to final wound healing. The purpose of this study was to quantify the levels of IL-10 in CVI ulcers and autologous donor tissue using Western blotting. Extracts were prepared from frozen wound tissue samples and equal amounts of protein were concentrated by immune-precipitation for Western blot analysis. Densitometric analysis was performed on nonsaturated chemilumigraphs and normalized to an IL-10 standard run on each gel. The quantity of IL-10 in CVI ulcers was found to be 490% of the quantity in autologous donor tissue. This study provides confirmatory quantitative data which supports previous immunohistochemical findings showing elevated levels of IL-10 in CVI ulcers.


Alteration of lipids and the transcription of lipid-related genes in myelodysplastic syndromes via a TP53-related pathway.
Myelodysplastic syndromes (MDS) are clonal stem cell diseases of the bone marrow characterized by abnormalities in maturation of hematopoietic cells of all lineages. MDS patients frequently have lower lipids and high rates of apoptosis and p53 (TP53) expression. An association between the reduced lipids in MDS and the expression of lipid-related genes was sought. We further evaluated whether 3-hydroxy-3-methyl-glutaryl-CoA reductase (HMGcoAR) and low-density lipoprotein receptor (LDL-R) are regulated by TP53 in vivo and in vitro. Gene expression was measured using real-time reverse transcription polymerase chain reaction on RNA extracted from bone marrow and peripheral blood from eight newly diagnosed MDS patients and eight controls and from mice livers. Serum lipid profile was measured using colorimetric enzymatic procedures. Total- and LDL cholesterol were lower in MDS patients in comparison to controls (p = 0.04 and p = 0.01, respectively). HMGcoAR messenger RNA increased in peripheral blood and bone marrow of MDS patients compared to controls (p = 0.04 and p = 0.01, respectively). LDL-R messenger RNA was higher only in the peripheral blood of MDS patients (p = 0.05). Comparable results were obtained in vivo. The transcription of these genes correlates with TP53 activation as documented by p21 messenger RNA elevation, a surrogate for TP53 activation and by using TP53 temperature-sensitive cells treated with adriamycin. To conclude, an association between reduced lipids in MDS and expression of HMGcoAR and LDL-R genes was documented. The transcription of these genes can be regulated by TP53.


Impulse control disorders, dopamine dysregulation syndrome and sex dysfunction in Parkinson's disease.
Hypersexuality (HS), characterized by an aberrant, compulsive and inappropriate sexual drive, is an underreported and undermanaged complication of the treatment with dopamine agonists in patients with Parkinson's disease (PD). HS is part of the spectrum of impulse control disorders (ICDs). The failure to control these addictive behaviors is distressing for the patient and it is associated with important consequences. Reports of the prevalence of HS showed different results, due to the lack of standardized diagnostic criteria, but also due to the embarrassment or guilt feelings that patients experience regarding their sexual behaviors. Overall, it is considered that HS may occur in 1.92-22.8% of PD patients and the main risk factors involved could be male sex and genetic susceptibility. The pathophysiology of HS is unknown, but several degenerative mechanisms were proposed, involving dopaminergic, serotoninergic and noradrenergic pathways in cerebral regions responsive for planning and rewarding. There are no standardized tools for the assessment of HS in PD patients; however, several scales and questionnaires were developed with the aim of screening and rating the severity of ICDs, including HS. The management of HS is challenging. Discontinuation or reducing the dopaminergic treatment was effective in some cases, but it might lead to worsening of the motor function. A multidisciplinary approach is mandatory to manage the other associated aspects, such as psychological and social consequences of HS.


Pten heterozygosity restores neuronal morphology in fragile X syndrome mice.
Genetic studies of hippocampal granule neuron development have been used to elucidate cellular functions of Pten and Fmr1. While mutations in each gene cause neurodevelopmental disorders such as autism and fragile X syndrome, how Pten and Fmr1 function alone or together during normal development is not known. Moreover, Pten mRNA is bound by the fragile X mental retardation protein (FMRP) RNA binding protein, but how this physical interaction impinges on phosphatase and tensin homolog protein (PTEN) expression is not known. To understand the interaction of PTEN and FMRP, we investigated the dentate gyrus granule neuron development in Pten and Fmr1 knockout (KO) mice. Interestingly, heterozygosity of Pten restored Fmr1 KO cellular phenotypes, including dendritic arborization, and spine density, while PTEN protein expression was significantly increased in Fmr1 KO animals. However, complete deletion of both Pten and Fmr1 resulted in a dramatic increase in dendritic length, spine density, and spine length. In addition, overexpression of PTEN in Fmr1 KO Pten heterozygous background reduced dendritic length, arborization, spine density, and spine length including pS6 levels. Our findings suggest that PTEN levels are negatively regulated by FMRP, and some Fmr1 KO phenotypes are caused by dysregulation of PTEN protein. These observations provide evidence for the genetic interaction of PTEN and FMRP and a possible mechanistic basis for the pathogenesis of Fmr1-related fragile X neurodevelopmental disorders.


[Corticosteroid hormones and histones (on the mechanism of action of hormones)].
The role of interaction between corticosteroid hormones and histones is considered with regard to hormonal activation of genes and some secondary changes of cells functional condition due to the presence of the hormones.


Frequency-dependent depression of Vmax of slow response action potentials by a high concentration of papaverine.
The maximum upstroke velocity (Vmax) of Ca2+-dependent action potentials was used to measure relative changes in the slow inward current (Isi) caused by 10(-4) M papaverine as a function of pacing frequency in K-depolarized guinea pig ventricular strips. Papaverine caused a pronounced frequency-dependent depression of Vmax presumed to be correlated with a decrease in the rate of recovery from inactivation of Isi. Papaverine (10(-4) M) tripled the time constants for the biexponential recovery from depression of Vmax (determined by a paired pulse protocol) relative to the control. Depression of Vmax by papaverine during a 1 Hz stimulus train preceded by a long rest period reached a new steady state within three depolarizations. Complete poststimulation recovery from the depression of Vmax was also relatively rapid, unlike that described for verapamil. Preparations exposed to Mn2+ (after pretreatment with Ba2+ to restore excitability) showed a depressed steady-state Vmax independent of frequency. It is concluded that the frequency-dependent blockers of Vmax (papaverine and verapamil) differ in their rates of association and dissociation with the slow inward channel.


Plasma cysteine/cystine redox couple disruption in animal models of temporal lobe epilepsy.
Currently the field of epilepsy lacks peripheral blood-based biomarkers that could predict the onset or progression of chronic seizures following an epileptogenic injury. Thiol/disulfide ratios have been shown to provide a sensitive means of assessing the systemic redox potential in tissue and plasma. In this study, we utilized a rapid, simple and reliable method for simultaneous determination of several thiol-containing amino acids in plasma using HPLC with electrochemical detection in kainic acid (KA) and pilocarpine rat models of epilepsy. In contrast to GSH and GSSG levels, the levels of cysteine (Cys) were decreased by 42% and 62% and cystine (Cyss) were increased by 46% and 23% in the plasma of KA- and pilocarpine-injected rats, respectively after 48h. In chronically epileptic rats, plasma cysteine was decreased by 40.4% and 37.7%, and plasma GSSG increased by 33.8% and 35.0% following KA and pilocarpine, respectively. Treatment of rats with a catalytic antioxidant, 60min after KA or pilocarpine significant attenuated the decrease of plasma Cys/Cyss ratios at the 48h time point in both models. These observations suggest that the decreased cysteine and ratio of Cys/Cyss in plasma could potentially serve as redox biomarkers in temporal lobe epilepsy.


[Activity index in rheumatic polymyalgias].
The activity of rheumatic polymyalgia (RP) was assessed using simplified activity index (RPAI). 76 patients with a valid diagnosis of RP were treated in the Rheumatological Department of the Territorial Clinical Hospital #1; the clinical picture of the disease and its activity were evaluated according to RPAI, and the effect of glucocorticoids (GC) on the index was assessed. All the subjects had a high RPAI at admission (32.8 +/- 0.8). After three weeks of GC therapy RPAI decreased significantly. The frequency of RP recurrence correlated with the absence of remission after lowering prednisolone dose. Knowing the clinical manifestations and course of RP, the diagnosis is not difficult. However, for adequate treatment of patients the degree of disease activity needs to be assessed during monitoring, after dose lowering, and after prednisolone is discontinued.


Influence of prenatal noise and music on the spatial memory and neurogenesis in the hippocampus of developing rats.
During the prenatal period, the development of individual is influenced by the environmental factors. In the present study, the influence of prenatal noise and music on the spatial memory and neurogenesis in the hippocampus of developing rats was investigated. The exposure to the noise during pregnancy caused growth retardation, decreased neurogenesis in the hippocampus, and impaired spatial learning ability in pups. The exposure to music during pregnancy, on the other hand, caused increased neurogenesis in the hippocampus and enhanced spatial learning ability in pups. The present study has shown the importance of the prenatal environmental conditions for the cognition and brain development.


Mechanistic insights into chloride-related heart failure progression according to the plasma volume status.
Two types of heart failure (HF) progression were recently proposed on the basis of an increased vs. non-increased serum chloride concentration. The applicability of this concept to real-world HF pathophysiology requires further investigation. The present study evaluated the mechanisms of HF progression to a different type according to changes in the estimated plasma volume status (ePVS). Data from 47 patients (32% men; 78.2 ± 9.7 years of age) with stable to worsening HF (37.5 ± 16 days) were analysed. Physical examination, standard blood tests, and b-type natriuretic peptide (BNP) measurements were conducted. The ePVS was calculated as follows: ePVS (dL/g) = [100 - haematocrit (%)]/[haemoglobin (g/dL)]. For the study subjects as a whole (n = 47), changes in the ePVS correlated positively with changes in the serum chloride concentration from stable to worsening HF (r = 0.398, P = 0.0056). When divided into two groups of worsening HF with an increased (n = 31) vs. non-increased serum chloride concentration (n = 16), no significant baseline differences in body weight, serum logBNP, or ePVS were detected between groups. Under worsening HF, the increase in body weight (2.34 ± 1.12 vs. 2.59 ± 1.56 kg, P = 0.57) and logBNP (0.39 ± 0.30 vs. 0.54 ± 0.31 pg/mL, P = 0.13) did not differ between groups, but the increase in the ePVS was smaller in the group with a non-increased serum chloride concentration compared with that with an increased serum chloride concentration (0.292 ± 0.49 vs. 0.653 ± 0.60 dL/g, P = 0.044). An increase in the %change in ePVS ≥ 10% was less common in patients with a non-increased chloride concentration (37% vs. 71%, P = 0.03). Patients with a non-increased serum chloride concentration had more HF signs (3.31 ± 0.79 vs. 2.65 ± 0.71, P = 0.005) and a higher incidence of pulmonary rales (63% vs. 16%, P = 0.0024) than those with an increased serum chloride concentration. According to the changes in the ePVS, HF progression may result from a difference between two HF types (i.e. increased vs. non-increased serum chloride concentration) in the cardiac reserve in response to a given cardiac burden by modulating plasma volume status via the possible tonicity potential of chloride.


Dynamics and mechanistic interpretations of nonribosomal peptide synthetase cyclization domains.
Ox-/thiazoline groups in nonribosomal peptides are formed by a variant of peptide-forming condensation domains called heterocyclization (Cy) domains and appear in a range of pharmaceutically important natural products and virulence factors. Recent cryo-EM, crystallographic, and NMR studies of Cy domains make it opportune to revisit outstanding questions regarding their molecular mechanisms. This review covers structural and dynamical findings about Cy domains that will inform future bioengineering efforts and our understanding of natural product synthesis.


Clinical profile and role of C-reactive protein and lactate dehydrogenase as prognostic marker and comparative efficacy of different pharmacotherapeutic agents in patients with cerebrovascular accident: An observational study.
The current study was conducted to determine the role of C-reactive protein (CRP) and lactate dehydrogenase (LDH) as prognostic-marker and outcomes of different pharmacotherapeutic agents among patients with cerebrovascular accident (CVA). A hospital-based observational study was conducted and patients with CVA admitted were included. Serum-CRP on admission correlated positively with modified Rankin score (mRS) (r = 0.9, P < 0.001) in ischemic stroke, whereas no correlation between serum LDH with mRS (r = 0.1, P = 0.5) was observed. Neither CRP nor LDH was helpful in predicting the outcome in hemorrhagic stroke. The use of Vitamin B12 was associated with favorable outcome in ischemic CVA cases and use of folic acid was associated with favorable outcome among hemorrhagic-CVA patients.


The vasopressin gene non-canonical Hogness box: effect on protein binding and promoter function.
Comparison of the promoter sequences of the genes encoding the neuropeptide hormone vasopressin from a number of organisms has revealed that they do not contain a classical Hogness box. In all vertebrate species examined, the canonical TATA box is replaced with a CATA sequence. We hypothesised that this conserved modified sequence may play a role in the regulation of vasopressin promoter activity. We used electrophoretic mobility shift assays to show that TATA and CATA sequences generate different complexes with SON nuclear proteins. Further, the transfection of wild-type (CATA) and mutated (TATA) VP promoter-reporter constructs into a heterologous cell line demonstrated a sequence-specific effect on transcriptional activity. The CATA sequence contributes to weaker promoter activity than a TATA box, but is able to interact with the upstream elements to increase the efficacy of an enhancer. The CATA box may thus be involved in the cell-specific and physiological regulation of the VP gene.


Could the different estrus periods significantly affect body temperature on clinical examination in bitches?
Clinical examination is essential to make a differential diagnosis list, and body temperature measurement is an indispensable part of the clinical examination. Throughout the estrus cycle, body temperature may fluctuate high enough to affect the clinical examination results. Aim of this study, whether there were any differences between the different estrus periods, body parts (rectal, intravaginal and corneal), and methods (contact thermography and infrared thermography) in regard to temperature in bitches. The animal material of this study consisted of 61 healthy, non-pregnant, non-sportive bitches (2.78 ± 0.22 mean years of age; mean weight 23.81 ± 1.61kg) of different breeds. Corneal temperature measurement results were found the least constant and significant differences were observed between different estrus periods except in the anestrus period. Rectal temperature (RT) measurement results were the most constant. However, in ANOVA analysis, significant differences were observed between the different estrus periods, and the difference between the anestrus and diestrus periods was very close to 0.5 °C. Although, in the Bland-Altman plot analysis, the agreement was observed only between rectal and intravaginal temperature (p: 0.572, r2: 0.70), the limits of agreement were not clinically acceptable. In conclusion, neither corneal temperature measurement with infrared technology nor intravaginal temperature measurement should not be an alternative to RT measurement, and RT measurement results should evaluate carefully considering the estrus period.


Genetic diversity of Rhodopirellula strains.
Rhodopirellula baltica SH1(T) is a marine planctomycete with 7,325 genes in its genome. Ten strains of the genus Rhodopirellula were studied in whole genome microarray experiments to assess the extent of their genetic relatedness to R. baltica SH1(T). DNA of strains which were previously affiliated with the species R. baltica (OTU A) hybridized with 3,645-5,728 genes of the type strain on the microarray. Strains SH398 and 6C (OTU B), representing a closely related species with an average nucleotide identity of 88 %, showed less hybridization signals: 1,816 and 3,302 genes gave a hybridization signal, respectively. Comparative genomics of eight permanent draft genomes revealed the presence of over 4,000 proteins common in R. baltica SH1(T) and strains of OTU A or B. The genus Rhodopirellula is characterized by large genomes, with over 7,000 genes per genome and a core genome of around 3000 genes. Individual Rhodopirellula strains have a large portion of strain-specific genes.


Excision repair of ultraviolet damage in mammalian cells. Evidence for two steps in the excision of pyrimidine dimers.
The incidence of pyrimidine dimer formation and the kinetics of DNA repair in African green monkey kidney CV-1 cells after ultraviolet (UV) irradiation were studied by measuring survival, T4 endonuclease V-sensitive sites, the fraction of pyrimidine dimers in acid-insoluble DNA as determined by thin layer chromatography (TLC), and repair replication. CV-1 cells exhibit a survival curve with extrapolation number n = 7.8 and Do = 2.5 J/m2. Pyrimidine dimers were lost from acid-insoluble DNA more slowly than endonuclease-sensitive sites were lost from or new bases were incorporated into high molecular weight DNA during the course of repair. Growth of CV-1 cultures in [3H]thymidine or X-irradiation (2 or 10 krads) 24 h before UV irradiation had no effect on repair replication induced by 25 J/m2 of UV. These results suggest that pyrimidine dimer excision measurements by TLC are probably unaffected by radiation from high levels of incorporated radionuclides. The endonuclease-sensitive site and TLC measurements can be reconciled by the assumption that pyrimidine dimers are excised from high molecular weight DNA in acid-insoluble oligonucleotides that are slowly degraded to acid-soluble fragments.


An in vitro and in vivo study: Valencene protects cartilage and alleviates the progression of osteoarthritis by anti-oxidative stress and anti-inflammatory effects.
Osteoarthritis (OA) is a heterogeneous disease involving the whole joint. The pathogenesis involves oxidative stress levels and chronic inflammation, and Valencene (VA) has excellent anti-inflammatory and antioxidant stress abilities. The objective was to study the effects of VA therapy on combating oxidative stress and to evaluate the protective effect of chondrocytes to alleviate the progression of OA. C57BL6J mouse chondrocytes were used as the primary cells in this study. Mouse chondrocytes were stimulated with IL-1β, and VA was administered in different concentrations. Reactive oxygen species (ROS) assay kits, western blotting, cellular immunofluorescence, and scanning microscopy were used to evaluate VA's antioxidant stress mechanism, anti-inflammatory effect, and cartilage protective ability. The mouse arthritis model constructed by destabilization of medial meniscus (DMM) was observed by micro-CT scan and histology after different treatments. We found that VA can reverse the rise of ROS under IL-1β, the degeneration of the cartilage extracellular matrix, and the production of inflammatory mediators. In terms of mechanism, VA activated NRF2/HO-1/NQO1 pathway, thus enhancing ROS clearance. The phosphorylation of IκBα is inhibited, which further reduces the downstream phosphorylation of P65 in nuclear factor-κB (NF-κB) signaling. In addition, VA inhibited mitogen-activated protein kinase (MAPK) signaling molecules P-JNK, P-ERK, and P-P38, inhibiting the production of inflammatory mediators and thus inhibiting Aggrecan and Collagen Type II (COL2)degeneration. In vivo, VA reduced DMM-induced osteophytes and spurs, suppressed subchondral bone destruction, and reduced articular cartilage erosion. Our study demonstrated that VA is an effective candidate for OA treatment.


Integrated Assessment of Chemical and Biological Recovery After Diversion and Treatment of Acid Mine Drainage in a Rocky Mountain Stream.
Responses of stream ecosystems to gradual reductions in metal loading following remediation or restoration activities have been well documented in the literature. However, much less is known about how these systems respond to the immediate or more rapid elimination of metal inputs. Construction of a water treatment plant on the North Fork of Clear Creek (NFCC; CO, USA), a US Environmental Protection Agency Superfund site, captured, diverted, and treated the two major point-source inputs of acid mine drainage (AMD) and provided an opportunity to investigate immediate improvements in water quality. We conducted a 9-year study that included intensive within- and among-year monitoring of receiving-stream chemistry and benthic communities before and after construction of the treatment plant. Results showed a 64%-86% decrease in metal concentrations within months at the most contaminated sites. Benthic communities responded with increased abundance and diversity, but downstream stations remained impaired relative to reference conditions, with significantly lower taxonomic richness represented by a few dominant taxa (i.e., Baetis sp., Hydropsyche sp., Simulium sp., Orthocladiinae). Elevated metal concentrations from apparent residual sources, and relatively high conductivity from contributing major ions not removed during the treatment process, are likely limiting downstream recovery. Our study demonstrates that direct AMD treatment can rapidly improve water quality and benefit aquatic life, but effectiveness is limited, in part, to the extent that inputs of metals are captured and treated. Consideration should also be given to the effects of elevated major ion concentrations from the treated effluent not removed during the lime treatment process. Continued chemical and biological monitoring will be needed to quantify the NFCC recovery trajectory and to inform future remediation strategies. Environ Toxicol Chem 2023;42:512-524. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.


Salpingitis associated with the presence of nonmedicated IUDs.
Fallopian tube tissue obtained at Pomeroy sterilization of 75 normally cycling women, 75 women 2 to 3 days postpartum, and 75 asymptomatic women who had used a nonmedicated IUD for at least a year was examined histologically. There was an increased frequency of tissue edema and congestion in the postpartum group only. The frequency and extent of acute inflammatory changes in the three groups was not significantly different.


An in vitro coculture system of human peripheral blood mononuclear cells with hepatocellular carcinoma-derived cells for predicting drug-induced liver injury.
Preventing clinical drug-induced liver injury (DILI) remains a major challenge, because DILI develops via multifactorial mechanisms. Immune and inflammatory reactions are considered important mechanisms of DILI; however, biomarkers from in vitro systems using immune cells have not been comprehensively studied. The aims of this study were (1) to identify promising biomarker genes for predicting DILI in an in vitro coculture model of peripheral blood mononuclear cells (PBMCs) with a human liver cell line, and (2) to evaluate these genes as predictors of DILI using a panel of drugs with different clinical DILI risk. Transcriptome-wide analysis of PBMCs cocultured with HepG2 or differentiated HepaRG cells that were treated with several drugs revealed an appropriate separation of DILI-positive and DILI-negative drugs, from which 12 putative biomarker genes were selected. To evaluate the predictive performance of these genes, PBMCs cocultured with HepG2 cells were exposed to 77 different drugs, and gene expression levels in PBMCs were determined. The MET proto-oncogene receptor tyrosine kinase (MET) showed the highest area under the receiver-operating characteristic curve (AUC) value of 0.81 among the 12 genes with a high sensitivity/specificity (85/66%). However, a stepwise logistic regression model using the 12 identified genes showed the highest AUC value of 0.94 with a high sensitivity/specificity (93/86%). Taken together, we established a coculture system using PBMCs and HepG2 cells and selected biomarkers that can predict DILI risk. The established model would be useful in detecting the DILI potential of compounds, in particular those that involve an immune mechanism.


Multidrug-resistant OXA-48/CTX-M-15 Klebsiella pneumoniae cluster in a COVID-19 intensive care unit: salient lessons for infection prevention and control during the COVID-19 pandemic.
Wards caring for COVID-19 patients, including intensive care units (ICUs), have an important focus on preventing transmission of SARS-CoV-2 to other patients and healthcare workers. To describe an outbreak of carbapenemase-producing Enterobacterales (CPE) in a COVID-19 ICU and to discuss key infection control measures enabling prompt termination of the cluster. CPE were isolated from clinical specimens and screening swabs from intensive care patients with COVID-19 disease and from environmental screening. Whole-genome sequencing analysis was instrumental in informing phylogenetic relationships. Seven clinical isolates and one environmental carbapenemase-producing Klebsiella pneumoniae isolate - all carrying OXA-48, CTX-M-15 and outer membrane porin mutations in ompK35/ompK36 - were identified with ≤1 single nucleotide polymorphism difference, indicative of clonality. A bundle of infection control interventions including careful adherence with contact precautions and hand hygiene, twice weekly screening for multidrug-resistant organisms, strict antimicrobial stewardship, and enhanced cleaning protocols promptly terminated the outbreak. Prolonged use of personal protective equipment is common with donning and doffing stations at the ward entrance, leaving healthcare workers prone to reduced hand hygiene practices between patients. Minimizing transmission of pathogens other than SARS-CoV-2 by careful adherence to normal contact precautions including hand hygiene, even during high patient contact manoeuvres, is critical to prevent outbreaks of multidrug-resistant organisms. Appropriate antimicrobial stewardship and screening for multidrug-resistant organisms must also be maintained throughout surge periods to prevent medium-term escalation in antimicrobial resistance rates. Whole-genome sequencing is highly informative for multidrug-resistant Enterobacterales surveillance strategies.